Additional file 1 of Phototropin2 3’UTR overlaps with the AT5G58150 gene encoding an inactive RLK kinase
Creators: name: Łabuz, Justyna
name: Banaś, Agnieszka Katarzyna
name: Zgłobicki, Piotr
name: Bażant, Aneta
name: Sztatelman, Olga
name: Giza, Aleksandra
name: Lasok, Hanna
name: Prochwicz, Aneta
name: Kozłowska-Mroczek, Anna
name: Jankowska, Urszula
name: Hermanowicz, Paweł
Publisher: name: figshare
Year: 2024
Subjects: subject: Genetics
subject: FOS: Biological sciences
subject: Plant Biology

description: Additional file 1: Fig. S1. The locus of PHOT2 (AT5G58140) and AT5G58140 genes in the whole-genome pairwise alignments between Arabidopsis and several plant genomes, obtained with the VISTA-Point tool ( https://pipeline.lbl.gov/ ). For each species, the curves show the identity score of the alignment, averaged across a 100 bp moving window. A region is considered to be conserved if the sequence identity is at least 70% over at least 100 bp. Conserved regions are color-coded as dark blue, light blue or orange if they correspond to exons, UTRs, or non-coding sequences, respectively. Fig. S2. Confirmation of at5g58150 mutation (SALK_093781C, insertion in the promoter region) with the three primers in one reaction (Lba1, sequence specific primers Table S1). Fig. S3. Kinase activity assay for AT5G58150 kinase domain. Fig. S4. Laser scanning confocal images of N. benthamiana epidermal cells transiently co-expressing AT5G58150-GFP or Plasma Membrane-mCherry or Tonoplast-mCherry markers. Fig. S5. Western Blot analysis of N. benthamiana epidermal cells transiently co-expressing PHOT2 and AT5G58150 fused with C(N)-terminal YFP fragments in the following configurations: AT5G58150_NtermYFP and AT5G58150_CtermYFP, NtermYFP_PHOT2 and PHOT2_CtermYFP, NtermYFP_PHOT2 and AT5G58150_CtermYFP, CtermYFP_PHOT2 and PHOT2_NtermYFP, PHOT2_CtermYFP and AT5G58150_NtermYFP, AT5G58150_NtermYFP, AT5G58150_CtermYFP, NtermYFP_PHOT2, CtermYFP_PHOT2, PHOT2_NtermYFP, PHOT2_CtermYFP probed with anti-cYFP and anti-nYFP. The white light image was merged with the chemiluminescent signal to show the borders of the membranes and molecular weight marker (PageRuler Prestained Protein, SM #26616, Thermo Scientific). Cropped image in Fig. 4. Fig. S6. AT5G58150 and phototropin2 interactions tested with MYTH assay. Fig. S7. Averaged curves (A) and amplitudes (B) of changes in rosette leaf transmittance T induced by blue light of increasing irradiance of 0.4, 1.6, 4, 20, 40, 80, 120 µmol·m-2·s-1 in wild type, at5g58150 mutant and 35S::AT5G58150-GFP lines. Fig. S8. Co-expression analysis of AT5G58150 based on proteomic data performed with Athena (https://athena.proteomics.wzw.tum.de/master_arabidopsisshiny/). Co-expression of AT5G58150 with BAK1, BSK, BRL1, BRL3 is observed. Fig. S9. Co-expression analysis of AT5G58150 based on transcriptomic data performed with Athena (https://athena.proteomics.wzw.tum.de/master_arabidopsisshiny/). Co-expression of AT5G58150 with ATBS1, BES1, BZR1 is observed. Table S6. List of proteins that co-immunoprecipitated with AT5G59150-GFP, identified with Mass Spectrometry, which fulfill the interaction criteria, together with the values of parameters taken into account in the calculations. Table S7. List of proteins co-immunoprecipitated with AT5G59150-GFP, identified with MS, with their functional descriptions from the String database. Table S8. Primer sequences used in this study. Table S9. Genes co-expressed with AT5G58150 based on published data from transcriptomic analysis performed with the Arabidopsis Co-expression Tool (https://www.michalopoulos.net/act). Genes of the brassinosteroid pathway, involved in root development as well as cell wall and junction formation, are marked in yellow.