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<article doi="10.1016/j.celrep.2013.06.034" abstract="A key feature in Huntington disease (HD) is the accumulation of mutant Huntingtin (HTT) protein, which may be regulated by posttranslational modifications. Here, we define the primary sites of SUMO modification in the amino-terminal domain of HTT, show modification downstream of this domain, and demonstrate that HTT is modified by the stress-inducible SUMO-2. A systematic study of E3 SUMO ligases demonstrates that PIAS1 is an E3 SUMO ligase for both HTT SUMO-1 and SUMO-2 modification and that reduction of dPIAS in a mutant HTT Drosophila model is protective. SUMO-2 modification regulates accumulation of insoluble HTT in HeLa cells in a manner that mimics proteasome inhibition and can be modulated by overexpression and acute knockdown of PIAS1. Finally, the accumulation of SUMO-2-modified proteins in the insoluble fraction of HD postmortem striata implicates SUMO-2 modification in the age-related pathogenic accumulation of mutant HTT and other cellular proteins that occurs during HD progression."><fig id="4998"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=4998"/><sd-panel panel_id="15219">(B) <sd-tag id="sdTag1" source="sdapi" category="time" entity_type="" role="" text="Time course" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Time course</sd-tag> of <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> modification of <sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag>-tagged <sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p (25Q)-purified proteins (WT, K6R, K9R, K6,9R, K6,9,15R, and S13,16D) was performed in vitro. SUMOylation was visualized using anti-<sd-tag id="sdTag6" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag> antibody. WT <sd-tag id="sdTag5" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p is SUMOylated within 16 min, K6R or K9R mutations delay SUMOylation, and combined mutations (K6,9R or K6,9,15R) greatly reduce SUMOylation. Mutations that mimic phosphorylation (S13,16D) alter kinetics of <sd-tag id="sdTag7" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> modification with SUMO modification observed beginning by 4 min.<graphic href="https://api.sourcedata.io/file.php?panel_id=15219"/></sd-panel></fig><fig id="4999"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=4999"/><sd-panel panel_id="15220">(A) <sd-tag id="sdTag10" source="sdapi" category="assay" entity_type="" role="" text="qRT-PCR" ext_ids="BAO_0002084" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="real-time PCR " ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">qRT-PCR</sd-tag> analysis of SUMO-modifying proteins and enzymes in <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="cortex" ext_ids="UBERON:0000956" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="cerebral cortex" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">cortex</sd-tag> and <sd-tag id="sdTag12" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="striata" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striata</sd-tag> of WT <sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> at 12 weeks. Relative expression for all the SUMO enzymes is normalized to <sd-tag id="sdTag15" source="sdapi" category="entity" entity_type="organism" role="component" text="mouse" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mouse</sd-tag> <sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="gene" role="normalizing" text="β-actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">β-actin</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15220"/></sd-panel><sd-panel panel_id="15221">(B and C) <sd-tag id="sdTag34" source="sdapi" category="assay" entity_type="" role="" text="qRT-PCR" ext_ids="BAO_0002084" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="real-time PCR " ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">qRT-PCR</sd-tag> of SUMO mRNAs from 12-week-old WT and <sd-tag id="sdTag26" source="sdapi" category="entity" entity_type="organism" role="experiment" text="R6/2" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">R6/2</sd-tag> <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="organism" role="experiment" text="mouse" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mouse</sd-tag> <sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="cortex" ext_ids="UBERON:0000956" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="cerebral cortex" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">cortex</sd-tag> (B) and <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="striatum" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striatum</sd-tag> (C). SUMO enzyme mRNAs are differentially expressed in <sd-tag id="sdTag37" source="sdapi" category="entity" entity_type="organism" role="experiment" text="R6/2" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">R6/2</sd-tag> versus control with statistically significant increases in <sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SENP1" ext_ids="223870" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Senp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP1</sd-tag> (p = 0.01), <sd-tag id="sdTag28" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SENP3" ext_ids="80886" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Senp3" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP3</sd-tag> (p = 0.02), and <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SUMO-1" ext_ids="22218" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Sumo1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> (p = 0.003) in <sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="cortex" ext_ids="UBERON:0000956" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="cerebral cortex" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">cortex</sd-tag> and <sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SENP1" ext_ids="223870" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Senp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP1</sd-tag> (p = 0.02), <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SENP6" ext_ids="215351" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Senp6" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP6</sd-tag> (p = 0.04), <sd-tag id="sdTag31" source="sdapi" category="entity" entity_type="gene" role="assayed" text="PIAS3" ext_ids="229615" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Pias3" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS3</sd-tag> (p = 0.007), <sd-tag id="sdTag32" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SUMO-1" ext_ids="22218" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Sumo1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> (p = 0.02), and <sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="gene" role="assayed" text="SUMO-2" ext_ids="170930" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Sumo2" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag> (p = 0.02) in <sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="striatum" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striatum</sd-tag>. Samples were analyzed in quadruplicate and normalized to <sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="organism" role="experiment" text="mouse" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mouse</sd-tag> <sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="gene" role="normalizing" text="β-actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">β-actin</sd-tag>. Data are shown as <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="organism" role="experiment" text="R6/2" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">R6/2</sd-tag> expression relative to WT levels set at 1 for each enzyme with ± SD (n = 4). *p < 0.05. n.s., not significant.<graphic href="https://api.sourcedata.io/file.php?panel_id=15221"/></sd-panel><sd-panel panel_id="15222">(D and E) <sd-tag id="sdTag49" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> analysis of <sd-tag id="sdTag52" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61957" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sumo2" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> in 14-week <sd-tag id="sdTag55" source="sdapi" category="entity" entity_type="organism" role="experiment" text="R6/2" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">R6/2</sd-tag> <sd-tag id="sdTag56" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="cortex" ext_ids="UBERON:0000956" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="cerebral cortex" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">cortex</sd-tag> (D) and <sd-tag id="sdTag57" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="striatum" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striatum</sd-tag> (E) versus aged-matched controls. <sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61957" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sumo2" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> is upregulated in <sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="organism" role="experiment" text="R6/2" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">R6/2</sd-tag> <sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="tissue" role="experiment" text="striatum" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striatum</sd-tag> versus control (p = 0.026; n = 4). Protein is normalized to <sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="α-tubulin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">α-tubulin</sd-tag> and quantitated using ImageJ.<graphic href="https://api.sourcedata.io/file.php?panel_id=15222"/></sd-panel></fig><fig id="5000"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5000"/><sd-panel panel_id="15223">(A) <sd-tag id="sdTag62" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells transfected with <sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag>-tagged <sd-tag id="sdTag63" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p (46Q) or 46QP-K6,9,15R (3R) along with <sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> or <sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag>, lysed under denaturing conditions, and <sd-tag id="sdTag115" source="sdapi" category="entity" entity_type="molecule" role="component" text="nickel" ext_ids="CHEBI:28112" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="nickel atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">nickel</sd-tag> purified (<sd-tag id="sdTag116" source="sdapi" category="entity" entity_type="undefined" role="component" text="Ni-NTA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ni-NTA</sd-tag>). Unmodified <sd-tag id="sdTag72" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-46Q is indicated by the arrow and SUMO-modified <sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> by the boxed region. The lysine mutant (3R) serves as a negative control. <sd-tag id="sdTag117" source="sdapi" category="entity" entity_type="undefined" role="component" text="Ni-NTA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ni-NTA</sd-tag> represents <sd-tag id="sdTag118" source="sdapi" category="entity" entity_type="molecule" role="component" text="nickel" ext_ids="CHEBI:28112" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="nickel atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">nickel</sd-tag>-purified <sd-tag id="sdTag74" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag>-tagged <sd-tag id="sdTag75" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>, and WC TCA represents 10% of the whole-cell lysate expression of <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> and <sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>-<sd-tag id="sdTag67" source="sdapi" category="entity" entity_type="protein" role="component" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> (transfection control). <sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> is modified by <sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> (left) and <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> (right).<graphic href="https://api.sourcedata.io/file.php?panel_id=15223"/></sd-panel><sd-panel panel_id="15224">(B) SUMO isopeptidases (<sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP1" ext_ids="29843" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP1</sd-tag>, <sd-tag id="sdTag81" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP2" ext_ids="59343" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP2</sd-tag>, <sd-tag id="sdTag84" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP3" ext_ids="26168" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP3" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP3</sd-tag>, <sd-tag id="sdTag82" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP5" ext_ids="205564" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP5</sd-tag>, and <sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP6" ext_ids="26054" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP6" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP6</sd-tag>) modulate SUMO-modified <sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> when overexpressed together with <sd-tag id="sdTag87" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (46QP-H4 or 3R) and <sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> (<sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag>). <sd-tag id="sdTag91" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP1" ext_ids="29843" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP1</sd-tag>, <sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP2" ext_ids="59343" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP2</sd-tag>, and <sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SENP6" ext_ids="26054" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SENP6" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SENP6</sd-tag> decrease <sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> SUMOylation. Graph depicts quantitation of <sd-tag id="sdTag83" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag> using the Odyssey Infrared Imaging System (LI-COR) to calculate the ratio of <sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> purified versus the <sd-tag id="sdTag96" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> modified by SUMO multiplied by 100.<graphic href="https://api.sourcedata.io/file.php?panel_id=15224"/></sd-panel><sd-panel panel_id="15225">(C) Titration of <sd-tag id="sdTag105" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag>. Denaturing <sd-tag id="sdTag119" source="sdapi" category="entity" entity_type="molecule" role="component" text="nickel" ext_ids="CHEBI:28112" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="nickel atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">nickel</sd-tag> purification of <sd-tag id="sdTag100" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p (46QP-H4) following transfection with decreasing amounts of <sd-tag id="sdTag101" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> reduces the amount of SUMO-modified <sd-tag id="sdTag106" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> to undetectable levels. The <sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="undefined" role="component" text="Ni-NTA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ni-NTA</sd-tag> blot displayed in the gray scale shows purified <sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1 and SUMO-modified <sd-tag id="sdTag103" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1 using <sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> antibody. Note that 0.5 μg of <sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> (¼ the amount of <sd-tag id="sdTag109" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> cDNA ) was used for identifying the <sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> E3 ligase for <sd-tag id="sdTag104" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p. Graph depicts quantitation of <sd-tag id="sdTag111" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag> using the Odyssey Infrared Imaging System to calculate the ratio of <sd-tag id="sdTag112" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> purified versus the <sd-tag id="sdTag113" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> modified by SUMO multiplied by 100. Note that all experiments were performed in triplicate, and a representative figure is shown.<graphic href="https://api.sourcedata.io/file.php?panel_id=15225"/></sd-panel></fig><fig id="5001"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5001"/><sd-panel panel_id="15226">(A) Under limiting SUMO conditions (1/4 <sd-tag id="sdTag124" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag>, lanes 3-9), <sd-tag id="sdTag123" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> increases <sd-tag id="sdTag125" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag>-SUMO modification above 1/4 SUMO alone. Purified <sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (arrow) and SUMO-<sd-tag id="sdTag127" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (boxed region) were detected using anti-<sd-tag id="sdTag128" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>. Graph depicts quantitation of the <sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="undefined" role="component" text="Ni-NTA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ni-NTA</sd-tag> <sd-tag id="sdTag130" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag> using the Odyssey Infrared Imaging Software (LI-COR) to calculate the ratio of <sd-tag id="sdTag131" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> purified versus the <sd-tag id="sdTag132" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> modified by SUMO multiplied by 100.<graphic href="https://api.sourcedata.io/file.php?panel_id=15226"/></sd-panel><sd-panel panel_id="15227">(B) <sd-tag id="sdTag140" source="sdapi" category="assay" entity_type="" role="" text="Western analysis" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Western analysis</sd-tag> of overexpression of <sd-tag id="sdTag141" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag>ex1 (46QP-H4 or 3R), <sd-tag id="sdTag142" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> (<sd-tag id="sdTag143" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag144" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag>), and all the PIAS proteins (<sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag>, <sd-tag id="sdTag155" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIASxα" ext_ids="9063" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIASxα</sd-tag>, <sd-tag id="sdTag156" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIASxβ" ext_ids="9063" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIASxβ</sd-tag>, <sd-tag id="sdTag146" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS3" ext_ids="10401" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS3" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS3</sd-tag>, and <sd-tag id="sdTag157" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIASy" ext_ids="51588" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS4" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIASy</sd-tag>). Under nonlimiting <sd-tag id="sdTag147" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> conditions, <sd-tag id="sdTag148" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> enhances SUMO modification of <sd-tag id="sdTag149" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>. WC TCA shows overall <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>-<sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> (transfection control) and <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> levels. Graph quantitating the ratio of <sd-tag id="sdTag153" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> purified versus the <sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> modified using the Odyssey (LICOR).<graphic href="https://api.sourcedata.io/file.php?panel_id=15227"/></sd-panel><sd-panel panel_id="15228">(C) Left panel is the <sd-tag id="sdTag160" source="sdapi" category="assay" entity_type="" role="" text="autoradiography" ext_ids="BAO_0002527" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="autoradiography" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">autoradiography</sd-tag> results of a <sd-tag id="sdTag161" source="sdapi" category="assay" entity_type="" role="" text="GST pull-down assay" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GST pull-down assay</sd-tag> showing that radiolabeled human <sd-tag id="sdTag162" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PIAS1" ext_ids="O75925" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PIAS1</sd-tag> interacts with <sd-tag id="sdTag163" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1p. Right panel is a phosphorimager analysis of <sd-tag id="sdTag166" source="sdapi" category="assay" entity_type="" role="" text="GST pull-downs" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GST pull-downs</sd-tag>, performed in triplicate, showing the percentage of <sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="35S" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">35S</sd-tag>-labeled PIAS protein that bound the <sd-tag id="sdTag167" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GST" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GST</sd-tag> proteins: <sd-tag id="sdTag168" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GST" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GST</sd-tag> alone, unexpanded <sd-tag id="sdTag169" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> with and without the proline-rich region (20QP and 20Q), expanded <sd-tag id="sdTag170" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> with and without the proline-rich region (51QP and 51Q), and the proline-rich region alone (Pro). Error bars were calculated as an estimate of SE = STDEV(n)/SQRT(n-1).<graphic href="https://api.sourcedata.io/file.php?panel_id=15228"/></sd-panel></fig><fig id="5002"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5002"/><sd-panel panel_id="15230">(B) Longer <sd-tag id="sdTag177" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> polypeptides are modified by <sd-tag id="sdTag178" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag>. <sd-tag id="sdTag179" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> overexpressing <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag>ex1p (46QP-H4 or 3R), unexpanded <sd-tag id="sdTag201" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586 fragment (25Q-586 or 25Q-3R-586 aa), and expanded <sd-tag id="sdTag180" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586 fragment (137Q-586 or 137Q-3R-586 aa) with <sd-tag id="sdTag174" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> (<sd-tag id="sdTag181" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag182" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag>). Cell lysates were subjected to <sd-tag id="sdTag183" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> <sd-tag id="sdTag175" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitation" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitation</sd-tag> (<sd-tag id="sdTag176" source="sdapi" category="assay" entity_type="" role="" text="IP" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">IP</sd-tag>) using <sd-tag id="sdTag184" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> antibody. <sd-tag id="sdTag185" source="sdapi" category="assay" entity_type="" role="" text="Western analysis" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Western analysis</sd-tag> performed with the Odyssey (LI-COR) allows detection of <sd-tag id="sdTag186" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (data not shown) and SUMO simultaneously and shows that all forms of <sd-tag id="sdTag187" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> are <sd-tag id="sdTag188" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> modified using the anti-<sd-tag id="sdTag189" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> antibody. HTTex1p is covalently <sd-tag id="sdTag190" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> modified (lane 3), and the modification disappears when Lys are mutated to Arg (3R) (lane 4). Both unexpanded (lane 4 and 5) and expanded <sd-tag id="sdTag191" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586 fragments (lane 6 and 7) are covalently <sd-tag id="sdTag192" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> modified in both the presence and absence of the three Lys in the N-terminal region of <sd-tag id="sdTag193" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (3R). Free <sd-tag id="sdTag194" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> is indicated with the arrow, and SUMO-modified <sd-tag id="sdTag195" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> is indicated by the boxes. Inset on the right, from a replicate experiment, shows comigration of expanded <sd-tag id="sdTag196" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (anti-<sd-tag id="sdTag197" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>) and <sd-tag id="sdTag198" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> (anti-<sd-tag id="sdTag199" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>) displayed in the gray scale and in color when the two antibodies are merged (<sd-tag id="sdTag200" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> in red, <sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-1" ext_ids="P63165" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-1</sd-tag> in green, and yellow when colocalizing).<graphic href="https://api.sourcedata.io/file.php?panel_id=15230"/></sd-panel><sd-panel panel_id="15231">(C) Bait plasmids (<sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586-25Q or <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586-73Q aa) were transformed into the <sd-tag id="sdTag204" source="sdapi" category="entity" entity_type="organism" role="component" text="L40ccua MATa" ext_ids="4932" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Saccharomyces cerevisiae" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L40ccua MATa</sd-tag> <sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="organism" role="component" text="yeast" ext_ids="4932" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Saccharomyces cerevisiae" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">yeast</sd-tag> strain. <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="organism" role="component" text="Yeast" ext_ids="4932" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Saccharomyces cerevisiae" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">Yeast</sd-tag> clones encoding bait proteins were individually mated against a matrix of MATα <sd-tag id="sdTag218" source="sdapi" category="entity" entity_type="organism" role="component" text="yeast" ext_ids="4932" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Saccharomyces cerevisiae" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">yeast</sd-tag> clones encoding 16,888 prey proteins (with <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="gene" role="component" text="Gal4" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Gal4</sd-tag> activation domain fusions) using pipetting and spotting robots. Diploid yeasts were spotted onto <sd-tag id="sdTag208" source="sdapi" category="entity" entity_type="undefined" role="component" text="SDIV" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDIV</sd-tag> (-<sd-tag id="sdTag209" source="sdapi" category="entity" entity_type="molecule" role="component" text="Leu" ext_ids="CHEBI:25017" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="leucine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Leu</sd-tag>-<sd-tag id="sdTag210" source="sdapi" category="entity" entity_type="molecule" role="component" text="Trp" ext_ids="CHEBI:27897" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="tryptophan" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Trp</sd-tag>-<sd-tag id="sdTag211" source="sdapi" category="entity" entity_type="molecule" role="component" text="Ura" ext_ids="CHEBI:17568" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="uracil" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Ura</sd-tag>-<sd-tag id="sdTag219" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag>) agar plates for selection of PPIs as well as nylon membranes placed on <sd-tag id="sdTag220" source="sdapi" category="entity" entity_type="undefined" role="component" text="SDIV" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDIV</sd-tag> <sd-tag id="sdTag221" source="sdapi" category="entity" entity_type="molecule" role="component" text="agar" ext_ids="CHEBI:2509" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="agar" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">agar</sd-tag> plates for <sd-tag id="sdTag214" source="sdapi" category="assay" entity_type="" role="" text="β-galactosidase" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">β-galactosidase</sd-tag> assays. After 5-6 days of incubation at 30ºC, digitized images of the agar plates and nylon membranes were assessed for growth and <sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="subcellular" role="reporter" text="β-galactosidase" ext_ids="GO:0009341" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="beta-galactosidase complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">β-galactosidase</sd-tag> activity using the software Visual Grid (GPC Biotech).<graphic href="https://api.sourcedata.io/file.php?panel_id=15231"/></sd-panel><sd-panel panel_id="15232">(D) Overexpression of <sd-tag id="sdTag225" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> alone, with unexpanded <sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> constructs (25Q-586 aa) plus <sd-tag id="sdTag224" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> (<sd-tag id="sdTag229" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag342" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag>), shows that <sd-tag id="sdTag227" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PIAS1" ext_ids="O75925" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PIAS1</sd-tag> binds both full-length <sd-tag id="sdTag230" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> and <sd-tag id="sdTag231" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>-586 fragment (25Q-586 aa). WT <sd-tag id="sdTag232" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> was used in these experiments to preclude confounding aggregation effects. Western analysis detection was performed using Odyssey and is displayed in the gray scale but is shown in color on the merge (<sd-tag id="sdTag234" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> is red, and <sd-tag id="sdTag228" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PIAS1" ext_ids="O75925" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PIAS1</sd-tag> is green). WB, <sd-tag id="sdTag235" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15232"/></sd-panel><sd-panel panel_id="15233">(E) <sd-tag id="sdTag240" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells overexpressing either expanded 586 aa-<sd-tag id="sdTag241" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> or the phosphomimetic (S13,16D = DD) with <sd-tag id="sdTag242" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> plus and minus <sd-tag id="sdTag243" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag>. <sd-tag id="sdTag249" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> was purified by <sd-tag id="sdTag245" source="sdapi" category="assay" entity_type="" role="" text="IP" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">IP</sd-tag> using <sd-tag id="sdTag343" source="sdapi" category="entity" entity_type="molecule" role="component" text="hydrazide" ext_ids="CHEBI:35362" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="hydrazide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">hydrazide</sd-tag> beads (Bioclone) crosslinked to <sd-tag id="sdTag246" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (Enzo) antibody and subjected to <sd-tag id="sdTag238" source="sdapi" category="assay" entity_type="" role="" text="western analysis" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">western analysis</sd-tag> using anti-<sd-tag id="sdTag247" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (<sd-tag id="sdTag258" source="sdapi" category="assay" entity_type="" role="" text="MAB5490" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MAB5490</sd-tag>). Arrows indicate <sd-tag id="sdTag239" source="sdapi" category="entity" entity_type="gene" role="component" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag>-modified <sd-tag id="sdTag248" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15233"/></sd-panel><sd-panel panel_id="15234">(F) <sd-tag id="sdTag251" source="sdapi" category="assay" entity_type="" role="" text="IP" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">IP</sd-tag> of <sd-tag id="sdTag252" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> with <sd-tag id="sdTag250" source="sdapi" category="entity" entity_type="molecule" role="component" text="hydrazide" ext_ids="CHEBI:35362" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="hydrazide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">hydrazide</sd-tag>-linked beads shows that both unexpanded and expanded <sd-tag id="sdTag253" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> 586 aa phosphomimetics (S13, 16D-586 aa) are modified by <sd-tag id="sdTag254" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag>. Arrows indicate SUMO-conjugated <sd-tag id="sdTag255" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>. Note that all experiments including the Y2H assay were done in triplicate; representative experiments are shown. Arrows indicate <sd-tag id="sdTag256" source="sdapi" category="entity" entity_type="gene" role="component" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag>-modified <sd-tag id="sdTag257" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15234"/></sd-panel></fig><fig id="5003"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5003"/><sd-panel panel_id="15235">(A) Western analysis of whole-cell lysates from <sd-tag id="sdTag268" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells transfected with <sd-tag id="sdTag269" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag>-<sd-tag id="sdTag262" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-1" ext_ids="7341" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-1</sd-tag> or <sd-tag id="sdTag270" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag> and/or 97Q-<sd-tag id="sdTag264" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> exon 1 and treated with 5 μM <sd-tag id="sdTag263" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="MG132" ext_ids="CHEBI:75142" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">MG132</sd-tag> for 18 hr. Lysates were separated using differential centrifugation into a detergent-soluble fraction (SOLUBLE) with 1% <sd-tag id="sdTag265" source="sdapi" category="entity" entity_type="molecule" role="component" text="Triton X-100" ext_ids="CHEBI:9750" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Triton X-100" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Triton X-100</sd-tag> and a detergent-insoluble fraction (INSOLUBLE) with 4% <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="molecule" role="component" text="SDS" ext_ids="CHEBI:8984" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sodium dodecyl sulfate" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">SDS</sd-tag>. <sd-tag id="sdTag271" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> probed with anti-<sd-tag id="sdTag272" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> shows full-length endogenous <sd-tag id="sdTag273" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> in the SOLUBLE fraction (upper arrow) and 97Q-<sd-tag id="sdTag267" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>ex1 (lower arrow) (left panel). In the INSOLUBLE fraction, <sd-tag id="sdTag274" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> HMW species are indicated by the bracket and asterisks (right panel).<graphic href="https://api.sourcedata.io/file.php?panel_id=15235"/></sd-panel><sd-panel panel_id="15236">(B) <sd-tag id="sdTag275" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="MG132" ext_ids="CHEBI:75142" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">MG132</sd-tag> and <sd-tag id="sdTag276" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag> cause mutant <sd-tag id="sdTag277" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> to accumulate as HMW species (bracke and asterisk). <sd-tag id="sdTag278" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> showing <sd-tag id="sdTag279" source="sdapi" category="assay" entity_type="" role="" text="IP" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">IP</sd-tag> with <sd-tag id="sdTag280" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> antibody crosslinked beads from the detergent-insoluble fraction probed with the anti-<sd-tag id="sdTag281" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=15236"/></sd-panel><sd-panel panel_id="15237">(C) Mutant <sd-tag id="sdTag285" source="sdapi" category="entity" entity_type="gene" role="intervention" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag> (97Q-Httex1) fibrils are detected with anti-<sd-tag id="sdTag286" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> in the insoluble fraction with treatment of <sd-tag id="sdTag287" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="MG132" ext_ids="CHEBI:75142" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">MG132</sd-tag> or addition of exogenous <sd-tag id="sdTag288" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15237"/></sd-panel><sd-panel panel_id="15238">(D) <sd-tag id="sdTag293" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> with increasing concentrations of <sd-tag id="sdTag294" source="sdapi" category="entity" entity_type="gene" role="intervention" text="SUMO-2" ext_ids="6613" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">SUMO-2</sd-tag> detected with anti-<sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="protein" role="reporter" text="His" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His</sd-tag> antibody (left panel). Middle panel is the same <sd-tag id="sdTag296" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag> probed with anti-<sd-tag id="sdTag297" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> showing soluble forms of <sd-tag id="sdTag298" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag>. Right panel presents <sd-tag id="sdTag299" source="sdapi" category="assay" entity_type="" role="" text="western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">western blot</sd-tag> from detergent-insoluble fraction with monomeric <sd-tag id="sdTag300" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (97Q) at 55 kDa, and the asterisk (*) indicates the HMW species. Note that all experiments were performed in triplicate; representative figures are shown.<graphic href="https://api.sourcedata.io/file.php?panel_id=15238"/></sd-panel></fig><fig id="5004"><title></title><label>Figure 7</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5004"/><sd-panel panel_id="15239">(A) <sd-tag id="sdTag304" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> analysis of <sd-tag id="sdTag305" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells over-expressing exogenous <sd-tag id="sdTag306" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> in the presence of mutant <sd-tag id="sdTag307" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (97Q) when separated into detergent-soluble and detergent-insoluble fractions. No difference is detected in monomeric <sd-tag id="sdTag308" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> (top panel, Soluble), but HMW <sd-tag id="sdTag309" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> levels increase with <sd-tag id="sdTag310" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> overexpression. Anti-<sd-tag id="sdTag303" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PIAS1" ext_ids="O75925" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PIAS1</sd-tag> antibody (Invitrogen) was used to detect <sd-tag id="sdTag311" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=15239"/></sd-panel><sd-panel panel_id="15240">(B) Acute knockdown of <sd-tag id="sdTag313" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> decreases HMW <sd-tag id="sdTag314" source="sdapi" category="entity" entity_type="protein" role="assayed" text="HTT" ext_ids="P42858" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">HTT</sd-tag> species in the detergent-insoluble fraction. <sd-tag id="sdTag312" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PIAS1" ext_ids="O75925" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PIAS1</sd-tag> knockdown is detected in detergent-soluble and -insoluble fractions using anti-<sd-tag id="sdTag315" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PIAS1" ext_ids="8554" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PIAS1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PIAS1</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=15240"/></sd-panel><sd-panel panel_id="15241">(C) <sd-tag id="sdTag318" source="sdapi" category="entity" entity_type="organism" role="assayed" text="Drosophila melanogaster" ext_ids="7227" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Drosophila melanogaster" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">Drosophila melanogaster</sd-tag> expressing mutant <sd-tag id="sdTag319" source="sdapi" category="entity" entity_type="gene" role="component" text="HTT" ext_ids="3064" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="HTT" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">HTT</sd-tag>ex1p (93Q) in a reduced <sd-tag id="sdTag322" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Su(var)2-10" ext_ids="35927" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Su(var)2-10" ext_tax_ids="7227" ext_tax_names="Drosophila melanogaster" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Su(var)2-10</sd-tag>/<sd-tag id="sdTag324" source="sdapi" category="entity" entity_type="gene" role="intervention" text="dPIAS" ext_ids="35927" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Su(var)2-10" ext_tax_ids="7227" ext_tax_names="Drosophila melanogaster" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">dPIAS</sd-tag> genetic background exhibits statistically significantly reduced <sd-tag id="sdTag321" source="sdapi" category="entity" entity_type="cell_type" role="assayed" text="photoreceptor neuron" ext_ids="CL:0000540///CL:0000210" norm_text="" ext_dbs="CL///CL" in_caption="True" ext_names="neuron///photoreceptor cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org//// https://identifiers.org/">photoreceptor neuron</sd-tag> degeneration (left panel, p = 0.033) when comparing <sd-tag id="sdTag323" source="sdapi" category="entity" entity_type="gene" role="intervention" text="dPIAS" ext_ids="35927" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Su(var)2-10" ext_tax_ids="7227" ext_tax_names="Drosophila melanogaster" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">dPIAS</sd-tag>/+ to +/+ <sd-tag id="sdTag326" source="sdapi" category="entity" entity_type="organism" role="assayed" text="flies" ext_ids="7227" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Drosophila melanogaster" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">flies</sd-tag> and increased overall <sd-tag id="sdTag344" source="sdapi" category="assay" entity_type="" role="" text="survival" ext_ids="OBI_0000699" norm_text="" ext_dbs="OBI" in_caption="True" ext_names="survival assessment" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ontobee.org/ontology/OBI?iri=http://purl.obolibrary.org/obo/">survival</sd-tag> (right panel, p = 0.047) when comparing <sd-tag id="sdTag328" source="sdapi" category="entity" entity_type="gene" role="intervention" text="dPIAS" ext_ids="35927" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Su(var)2-10" ext_tax_ids="7227" ext_tax_names="Drosophila melanogaster" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">dPIAS</sd-tag>1/+ to +/+ <sd-tag id="sdTag332" source="sdapi" category="entity" entity_type="organism" role="assayed" text="flies" ext_ids="7227" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Drosophila melanogaster" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">flies</sd-tag>. Significance was measured by Student's t test.<graphic href="https://api.sourcedata.io/file.php?panel_id=15241"/></sd-panel><sd-panel panel_id="15242">(D) HMW <sd-tag id="sdTag334" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SUMO-2" ext_ids="P61956" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUMO2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">SUMO-2</sd-tag> accumulates in postmortem HD <sd-tag id="sdTag335" source="sdapi" category="entity" entity_type="tissue" role="component" text="striata" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striata</sd-tag>. <sd-tag id="sdTag336" source="sdapi" category="assay" entity_type="" role="" text="Western blot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Western blot</sd-tag> analysis of the insoluble fraction from three control and three HD postmortem <sd-tag id="sdTag337" source="sdapi" category="entity" entity_type="tissue" role="component" text="striata" ext_ids="UBERON:0002435" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="striatum" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">striata</sd-tag> as described (Experimental Procedures).<graphic href="https://api.sourcedata.io/file.php?panel_id=15242"/></sd-panel></fig></article>