EMBO/sd-character-level-ner huggingface dataset:
a) A549 cells transfected with a GFP–Rubicon expression plasmid and were subjected to immunofluorescence with various organelle marker antibodies.figf7(a) A549 cells transfected with a GFP-Rubicon expression plasmid and were subjected to immunofluorescence with various organelle marker antibodies. Scale bar, 10 μm. (b) A549 cells stably expressing GFP-LC3 were subjected to Rubicon knockdown (KD) and then transfected with mStrawberry, mStrawberry-Rubicon or mStrawberry-Rubicon mutant lacking the Beclin 1 binding region (Δ393849). Cells with strong expression of mStrawberry-Rubicon showed an overexpression-dependent dominant-negative phenotype; therefore, we selected cells with modest expression. The number of GFP-LC3 puncta was counted and mean ± s.d. are shown. mStrawberry, n = 12 (12 cells); mStrawberry-Mm-Rubicon, n = 14 (14 cells); mStrawberry-Mm-RubiconΔ393-849, n = 13 (13 cells).
Rapamycin-induced autophagy is Ca2+-dependent.Figure 5 2+-dependent. Western-blot analysis for GAPDH and LC3 of protein lysates obtained from HeLa cells treated for 5 h with DMSO, 1 µM rapamycin (Rapa), 10 µM BAPTA-AM or both. One hour before harvesting, 100 nM bafilomycin A1 was added. Left: representative Western blots; right: quantification of the LC3-II/GAPDH ratio (n = 6). * p<0.05, repeated measurements ANOVA.
Figure 2WT mice (n=3-5) were treated with PBS or 12.5 mg/kg LPS with or without 10 mg/kg DEX pre-treatment. Ileum was isolated 6 hours after LPS challenge. (A) p-MLKL and MLKL protein levels and (B) RIPK3 protein levels were analyzed via western blot using ACTIN as a loading control. Expression levels were quantified using FIJI and normalized to ACTIN levels. P-values were calculated using 1-way ANOVA. Gene expression (Defa6, lysozyme) was measured via qPCR. P-values were calculated using 1-way ANOVA. All bars represent mean ± SEM. ****P< 0.0001; *** P<0.001; ** P≤ 0.01; * P≤ 0.05. H&E staining was performed on ileum samples. Scale bar: 50µm. Lysozyme expression (in red) was detected via IHC, with Hoechst counterstaining. Scale bars: 50µm. Quantification of lysozyme expression. For every picture (n=3-4 per treatment), a selection of 10 crypts was made. Lysozyme positive signal was quantified in this region of interest (ROI) using an automated script in QuPath. Values are the positive area relative to total area of ROI.
Figure 7Anti-hTAPBPL mAb (clone 54) neutralized the inhibitory activity of hTAPBPL-Ig on T cell proliferation Significance in was calculated by one‐way ANOVA with Dunnett test and others by two‐tailed Student's t‐test. * P<0.05, anti-hTAPBPL Ab group was compared with isotype Ab group. Anti-hTAPBPL mAb (clone 54) neutralized the inhibitory activity of hTAPBPL-Ig on CD69 expression by CD4+ and CD8+ T cells in vitro. Significance in was calculated by one‐way ANOVA with Dunnett test and others by two‐tailed Student's t‐test. * P<0.05, anti-hTAPBPL Ab group was compared with isotype Ab group. DBA/2J mice were injected s.c. with 1× 105 P388 murine leukemia cells, followed by injection of the anti-hTAPBPL mAb (25, 50, or 100µg) or isotype Ab (25, 50, or 100 µg) 3 times per week. Tumors size was measured over time. The mean tumor diameter (mm3) + S.D. at the indicated time points are shown. DBA/2J mice were injected s.c. with 1× 105 P388 murine leukemia cells, followed by injection of the anti-hTAPBPL mAb (25, 50, or 100µg) or isotype Ab (25, 50, or 100 µg) 3 times per week. At the end of the studies, tumors from 100 µg anti-hTAPBPL mAb- or isotype Ab-treated mice were harvested. Single-cell suspension of the tumors was analyzed for the percentage of (E, F) CD4+ T cells, CD8+ T cells, and (I and J) CD4+CD25+FoxP3+ Tregs by flow cytometry. Tumor infiltrating CD4+ and CD8+ T cells were examined by immunofluorescence. C57BL/6 mice were injected s.c. with 1x105 B16F10 melanoma cells, and intratumorally with anti-hTAPBPL mAb or isotype Ab (50 µg) 3 times per week. Tumors size was measured over time. The data are expressed as mean + SD and representative of 2 independent experiments with similar results (n=6/group/time).
Figure 8A) Volcano plot from RNA-seq analyses of Study 1 gastrocnemius muscles for tumor-bearing combination-treated mice versus tumor-bearing controls. Genes not differentially expressed in this comparison are indicated with blue. The remaining genes (red and green) are DEGs in the combination-treated versus tumor-bearing control comparison. The green coloring indicates the subset of these DEGs that are not also differentially expressed in the comparison of AR-42-treated tumor-bearing mice versus tumor-bearing controls, suggesting these genes are responsive to only the combination therapy. Log2-transformed fold change (FC) in expression is plotted on the x-axis and -log10 transformed Benjamini-Hochburg adjusted p-values are plotted on the y-axis. B) Significance values from transcription factor pathway-focused GSEA of GTx-024-treated tumor-free versus tumor-free control transcriptomes. *FDR< 1e-5 was determined for the CTNNB1 gene set, and was set to 1e-5 for the plot. C) Significance values from GSEA using combined CTNNB1 gene sets. Each treatment group was compared to tumor-free control transcriptomes. *FDR < 1e-5 determined, set to 1e-5 for plot. D) Enrichment plots from GSEA of the CTNNB1 gene set for each treatment group versus tumor-free control comparisons. GTx-024-treated tumor-free (blue checkered), tumor-bearing control (black), GTx-024-treated tumor-bearing (blue), AR-42-treated tumor-bearing (red) and Combination-treated tumor-bearing (green). E) mRNA expression of WNT effectors upstream of β-catenin. Data are presented as mean±SD of log-transformed fold change (log2FC) values versus tumor-free controls. Groups: Tumor-free control (n=6), tumor-free GTx-024 treated (n=5), tumor-bearing receiving vehicle (n=4), GTx-024 (n=5), AR-42 (n=5) or Combination (n=6).
The minor conformation of phosphoUbLong‐range HNCO experiments showing differences of hydrogen bonding patterns for major and minor phosphoUb species.Figure 4A Long‐range HNCO NMR experiments showing differences of hydrogen bonding patterns for major and minor phosphoUb species. Through H bond correlations (blue) recorded for residue Ile44 in the major and minor species of phosphoUb with reference to a control HNCO spectrum (red). In the major form of phosphoUb (left), the HN of Ile44 is a H bond donor to His68, as seen in wt Ub. In the minor form of phosphoUb (right), the HN of Ile44 does not have a cross peak to His68 but instead has a correlation to Val70. B Comparison of hydrogen bonding patterns for the β‐sheet portion of wt Ub [left, according to Nisius and Grzesiek ()], the major form of phosphoUb (middle) and the minor form of phosphoUb (right). Four Leu residues (green) undergo a two‐residue strand slippage to satisfy the observed hydrogen bonding pattern in the minor species. The green arrows indicate the resonances exemplarily shown in (A). C Structure of the major form of phosphoUb, where phosphoUb residues 1-61 are shown under a surface, and the β5 strand is shown in stick representation with red oxygen, blue nitrogen and purple phosphorus atoms. The four core Leu residues are shown in green.D Model of the phosphoUb minor species as in (C) in which the β5 strand is retracted into the protein core by two residues. This extends the β4-β5 loop and significantly shortens the C‐terminal tail of Ub.
Binding of TAB2 NZF to phosphoUb chainsStructure of TAB2 NZF (red) in complex with Lys63 diUb (cyan) [pdb‐id 2wwz (Kulathu et al, )].Figure 7A Structure of TAB2 NZF (red) in complex with Lys63 diUb (cyan) [pdb‐id 2wwz (Kulathu et al, )]. Ser65 for both Ub moieties is shown as spheres and highlighted with a blue circle. B Pull‐down assays with the Lys63‐specific TAB2 NZF against different tetraUb (Ub4) species are shown. Phosphorylation of K63 Ub4 does not prevent binding to TAB2 NZF. Met1‐linked tetraUb does not interact with TAB2 NZF, independent of Ser65 phosphorylation.
Figure 6Body weight in Flox and GRBATKO after 24 weeks of high-fat feeding (n = 10-13 animals per group). BAT and iWAT weight in Flox and GRBATKO mice fed HFD for 24 weeks (n = 10-13 animals per group). GR levels in BAT after 24 weeks high-fat feeding (n=10-13 mice per group). Representative H&E stained sections of BAT from Flox and GRBATKO animals. Scale bar 100 µm. Triglyceride content in BAT per gram of wet tissue (n = 10-13 animals per group). qRT-PCR analysis of thermogenic marker gene expression in BAT of Flox and GRBATKO mice after high-fat feeding (n = 9-13 animals per group). Representative immunohistochemistry for UCP1 in BAT. Scale bar 100 µm. Serum insulin levels determined by ELISA (n = 10-13 animals per group). Blood glucose in Flox and GRBATKO animals after 24 weeks HFD (n = 10-13 animals per group).
Figure 5(A&B) Cued and Contextual Fear-conditioning tests. Data represent the mean ± SEM of n = 6-8 mice per group; **P=0.0067 (3xTg+DSS vs 3xTg ctrl), **P=0.0059 (3xTg+DSS vs 3xTg R-vagotomy+DSS) (**P< 0.01), ***P=0.0009 (***P < 0.001), two-way ANOVA (A); **P=0.0086 (3xTg+DSS vs 3xTg ctrl), **P=0.0085 (3xTg+DSS vs 3xTg R-vagotomy+DSS) (**P< 0.01), ***P=0.0005 (***P < 0.001), one-way ANOVA (B). (C) Body weight changes of the experimental mice during the experimental paradigm. Data represent the mean± SEM of n=8 mice per group; ****P < 0.0001, two-way ANOVA. (D) Survival rate curve of the experimental mice during the experimental paradigm.
Figure 5A. Complex structures of the human β5c/β6 (left) and β5i/β6 (right) chimeric proteasomes with PR-924. The ligand binds in a linear (l) manner to the β5c/β6 substrate binding channel, but in a kinked (k) conformation to the β5i/β6 active site. The 2FO-FC electron density maps for the ligands bound to Thr1 of the chimeric β5 active sites are shown as blue meshes contoured at 1σ. The inhibitor and Thr1 have been omitted for phasing. B. Comparison of the conformations of ONX 0914, PR-924, 14, 16, 17 and 18 bound to the chimeric hβ5/hβ6 substrate binding channels. The l amino acid compounds ONX 0914 and 14 display a linear (l) binding mode to the hβ5c/hβ6 (left) and the hβ5i/hβ6 (right) chimeras, whereas the ligands 16 and 17 (P3-d-Ala and morpholine cap) adopt a kinked (k) conformation in both β5 substrate binding channels. Remarkably, the orientation of PR-924 and its analogue 18 (P3-d-Ala and 3-methyl-1H-indene cap) in the hβ5c/hβ6 chimera (and in WT yβ5; see Appendix Figure S5)) differs from that observed with the hβ5i/hβ6 chimera. Note that the inhibitors are rotated by about 45 ° compared to panel A.
Figure 6A A schematic diagram of the 4HRE-d2GFP fluorescent hypoxia reporter system containing a destabilized GFP (d2GFP) fluorescent reporter under the control of four repetitive cis-acting hypoxia-responsive elements (HRE) followed by a minimum CMV promoter. The hypoxia reporter system was constructed on the backbone of pCDH-CMV-MCS-EF1α-Neo lentiviral vector (System Biosciences). B Representative images of autonomous fluorescent signals from T387 and T3359 GSCs transduced with 4HRE-d2GFP in response to 5% oxygen. Strong GFP signals were detected under 5% oxygen, whereas no GFP signal was detected under 20% oxygen. Scale bar, 200 μm. C, Representative images of autonomous fluorescent signals of 4HRE-d2GFP in xenografts derived from or shUSP33- or shNT- expressing T387 GSCs transduced with 4HRE-d2GFP. Scale bar, 200 μm. , D The reduced fraction of GFP+ tumor cells in the xenografts derived from shUPS33-expressing GSCs relative to the control xenografts indicated that USP33 disruption impaired hypoxia response in tumor cells. ***, p < 0.001 (n = 5 tumors; mean ± s.e.m.; two tailed unpaired t-test). E, Representative images of autonomous fluorescent signals of 4HRE-d2GFP (green) and immunofluorescent staining of the stem cell marker SOX2 (red) in xenografts derived from T387 GSCs transduced with 4HRE-d2GFP. More GFP signals were detected in the SOX2+ cells compared with the SOX2- cells, suggesting a stronger hypoxia response in GSCs relative to NSTCs. Scale bar, 200 μm. More GFP signals were detected in the SOX2+ cells compared with the SOX2- cells, suggesting a stronger hypoxia response in GSCs relative to NSTCs. ***, p < 0.001 (n = 5 tumors; mean ± s.e.m.; two tailed unpaired t-test). G, Representative images of immunofluorescent staining of the apoptotic marker cleaved caspase 3 (red) and autonomous fluorescent signals of 4HRE-d2GFP (green) in xenografts derived from T387 GSCs transduced with 4HRE-d2GFP. The GFP+ population had much less apoptosis relative to the GFP- population, suggesting that hypoxia response inhibited cell apoptosis. Scale bar, 200 μm. The GFP+ population had much less apoptosis relative to the GFP- population, suggesting that hypoxia response inhibited cell apoptosis. (n = 5 tumors for each group; ***, p < 0.001; mean ± s.e.m.; two tailed unpaired t-test).
SUMO conjugation to RPA is dependent on Siz2 SAP domain and partially dependent on Rfa2 WH domain.Figure 4SUMO conjugation assays performed under multiple turnover conditions using RPA complexes with or without the WH domain of Rfa2 in presence of Siz2∆CT lacking (A) or including (B) its SAP domain. In both cases, reactions were done in technical triplicate and SDS-PAGE gels were stained with SYPRO. Only one representative gel is shown. Bands for Mw marker are fully annotated in Figure 1B. Histograms derived from the gels presented in (B) and presenting Rfa1 SUMO conjugation rates for RPA complexes with or without the WH domain of Rfa2. Histograms derived from the curves presented in Appendix Fig S5 and presenting apparent dissociation constants (Kd) of Siz2∆CT with DNA and its complexes with RPA or RPA∆WH.
Ligand-macromolecular docking analysis implicates species-specific GM3 recognition by TLR4/MD-2.Figure 9(A, B) Docking model of GM3 24:0 (A) and 16:0 (B) binding to human TLR4/MD-2 complex (3FXI). Basic residues of TLR4 are colored in blue. (C-E) Superposition of GM3 24:0 (in docking model) vs. Ra-LPS (in 3FXI) (C), GM3 24:0 vs. GM3 16:0 (in docking model) (D), and GM3 16:0 vs. lipid-IVa (in 2E59) (E). Basic residues and the dimer interface are indicated schematically. (F, G) Working model for hTLR4 activation enhanced by VLCFA-GM3 species (F) and reduced by LCFA-GM3 (G). Basic residues contributing to GM3 recognition are colored in blue. Residues of dimer interface are colored in red.
Figure 1A Scheme of sedimentation assay (created in BioRender.com): Phase separation of TDP-43 was induced by TEV protease cleavage of TDP-43-MBP-His6, and condensates were pelleted by centrifugation. B Sedimentation assay to quantify condensation of unmodified TDP-43 versus in vitro phosphorylated TDP-43 (+CK1δ, +ATP) and controls (CK1δ or ATP only); TDP-43 detected by Western blot (rabbit anti-TDP-43 N-term). Due to incomplete TEV cleavage, some TDP-43-MBP-His6 remains present and co-fractionates with cleaved TDP-43, due to TDP-43 self-self interaction. C Quantification of band intensities of cleaved TDP-43 shown as means of Supernatant/(Supernatant+Condensate) [S/(S+C)] ratio of three independent experimental replicates (n=3) ± SD. ***p < 0.0002 by one-way ANOVA with Dunnett´s multiple comparison test to Wt. D Schematic diagram of TDP-43 and sequence of C-terminal region (aa. 370 - 414) for Wt, phosphomimetic (S-to-D) variants and control (S-to-A) variants. NTD = N-terminal domain; RRM = RNA recognition motif, LCD = low complexity domain with α-helical structure. E Turbidity measurements (optical density (OD) at 600 nm) to quantify phase separation of the indicated TDP-43 variants at three different concentrations (in Hepes buffer). Values represent means of three independent experimental replicates (n=3) ± SD. *p < 0.0332, **p < 0.0021 and ***p < 0.0002 by one-way ANOVA with Dunnett´s multiple comparison test to Wt, comparing the respective concentration condition (5, 10, 20 µM). Representative bright field microscopic images of TDP-43 condensates (in Hepes buffer), Bar, 25 µm (F) and quantification of condensate number (G) Box plots show the comparison of median and inter-quartile range (upper and lower quartiles) of all fields of view (FOV) from Min to Max (whiskers) of two replicates (≥ 22 FOV per condition). *p < 0.0332, **p < 0.0021 and ***p < 0.0002 by one-way ANOVA with Dunnett´s multiple comparison test to Wt, comparing the respective concentration condition (5, 10, 20 µM). size (H) and roundness (I). Box plots show the comparison of median and inter-quartile range (upper and lower quartiles) of all fields of view (FOV) from Min to Max (whiskers) of two replicates (≥ 22 FOV per condition). *p < 0.0332, **p < 0.0021 and ***p < 0.0002 by one-way ANOVA with Dunnett´s multiple comparison test to Wt, comparing the respective concentration condition (5, 10, 20 µM).
Figure 2(A) Representative confocal images showing ChFP-α-syn transfected cells (in red) cultured alone (upper) or co-cultured with donor cells containing fluorescent Alexa-488 α-synuclein fibrils (in green) for 72 h (bottom). The arrow is pointing out representative α-synuclein fibrils Alexa-488 co-localized with ChFP-α-syn puncta. Scale bar represents 10 μm. Nuclei are stained with DAPI (blue) (n=3 independent experiments).(B) Quantification of the number of ChFP-α-synuclein puncta in ChFP-α-syn acceptor cells showed a significant increase in ChFP-α-synuclein puncta number when cells were co-cultured with donor cells containing α-synuclein fibrils (***, p < 0.001 by two-tailed Mann Whitney test).(C) Quantification of the percentage of co-localization of α-synuclein fibrils Alexa-488 and ChFP-α-syn puncta. Data shows mean ± s.e.m. After co-culture, 11% of the transferred α-synuclein fibrils Alexa-488 co-localized with ChFP-α-syn puncta
Figure 5Scheme showing the reaction catalyzed by Lro1. Generation of suppressors of choline auxotrophy of cho2opi3, cho2opi3lro1, and cho2opi3lro1 pLRO1 on choline-free medium as indicated at 30 oC for 7 d. Three suppressor clones derived of cho2opi3 and cho2opi3lro1 transformed with plasmid pLRO1 were spotted on SGal ura- plates without choline, incubated at 30 oC for 4 d, replica-plated onto SGal C+ and C- plates containing 5-FOA and uracil, and incubated at 30 oC for 7 d. Serial dilutions of the strains indicated were spotted on SD C+/- and incubated at 30 oC for 6 d. Growth (30 oC for 6 d) and phospholipid composition of co acc1N/H lro1 pLRO1 cultured in SD C- ura-, containing glucose/galactose mixtures (2%, w/v) as carbon source with the percentage of galactose indicated. Phospholipid composition analyzed by TLC is presented as mean percentage of total phospholipid of 3 biological replicates with the individual values indicated.
Figure 4Effects of Cyclo (A-C, n=14) on the progression of kindling including seizure class (A,D,G,J), evoked electrographic seizure duration (ESD) (B,E,H,K) and the number of stimulations required to reach equivalent seizure intensity (C,F,I,L). In A-F, Ctrl are vehicle-treated mice. Effects of 5E1 (D-F, n=16-20), on the progression of kindling including seizure class (A,D,G,J), evoked electrographic seizure duration (ESD) (B,E,H,K) and the number of stimulations required to reach equivalent seizure intensity (C,F,I,L). In A-F, Ctrl are vehicle-treated mice. Effects of ablation of Smo in CaMKIIα-positive neurons (G-I), on the progression of kindling including seizure class (A,D,G,J), evoked electrographic seizure duration (ESD) (B,E,H,K) and the number of stimulations required to reach equivalent seizure intensity (C,F,I,L). In G-I, Ctrl: Smofl/fl induced by tamoxifen; CaMK: Smofl/flCaMKIIα-CreERT2 induced by tamoxifen, n=14-20. Effects of ablation of Smo in Aldh1l1-positive astrocytes (J-L) on the progression of kindling including seizure class (A,D,G,J), evoked electrographic seizure duration (ESD) (B,E,H,K) and the number of stimulations required to reach equivalent seizure intensity (C,F,I,L). In J-L, Ctrl: Smo+/fl; Aldh1l1: Smo+/flAldh1l1-Cre, n=19-20. (M) Frequency of spontaneous seizures (class 4-5) in mice administrated with Cyclo or vehicle (Ctrl) at 4th, 6th and 8th week after pilocarpine SE induction. n=22-23. (N) Percentage of mice with class 5 seizures within 8 weeks after pilocarpine SE induction. n=22-23. Cyclo: 10 mg/kg. 5E1: 900 ng/mouse. Data were mean ± SEM from at least three independent experiments. *p<0.05; **p<0.01; ***p<0.001 vs. Ctrl with student's t test. (O) Schematic diagram depicting a working model for Shh-regulation of epileptogenesis. Epileptic activity triggers sequential responses, including Shh release, inhibition of glutamate transporter activity, and increase in extracellular glutamate, leading to epilepsy development.
Figure 5A) Domain organization of ADAM17 and conservation of the PP2A-B56 binding motif. B) KD values obtained by ITC measurements with full-length recombinant B56α and indicated ADAM17 variant peptides. C) The indicated full-length murine Myc-ADAM17 derivatives were transfected into HeLa cells stably expressing YFP-B56α. YFP-B56α was purified (IP) and ADAM17 binding determined by Western blotting. PP2A-C; catalytic subunit, PP2A-A; scaffold subunit. D) Dephosphorylation by the PP2A-B56α holoenzyme complex of the indicated phosphorylated GST-ADAM17 (V724-C827) fragments. The GST-ADAM17 (V724-C827) substrates was phosphorylated with radioactive ATP using Protein Kinase A and incubated with the PP2A-B56α holoenzyme. Removal of radioactive phosphate was monitored over time. The mean and Standard deviation of 4 independent experiments are shown. E) Protein expression of ADAM17 variants in the DLD-1 Adam17-/- cell line, determined by Western blot. GAPDH was used as an internal loading control. F) Amphiregulin (AREG) shedding measured by ELISA of conditioned media from untreated, H2O2 treated or irradiated with x-ray DLD-1 Adam17-/- cells (clone #1) expressing full-length ADAM17 variants (wt, I762A or LEE). Two sided, unpaired Students t-test test was applied to test for significant differences *p<0.05, **p<0.01, ***p<0.001. Mean and standard deviation indicated from at least three independent experiments. G) Exogenous wt and I762A full length ADAM17 was immuno-purified from Adam17-/- cells treated with H2O2 and subjected to label-free LC-MS/MS to determine differential phosphorylation status of T735 and S808 (S811 in murine ADAM17).
Figure 1(A) Phylogenetic tree of GNAT candidates from Arabidopsis thaliana (black letters), Saccharomyces cerevisiae (orange letters), and Escherichia coli (green letters) containing the acetyltransferase Pfam domains (PF0058, PF13302, PF13508, PF13673) GNAT family sequences were aligned with ClustalW, and a phylogenetic tree was designed by applying the Neighbor-Joining method. Bootstrap analysis was performed using 2000 replicates, whereby the resulting bootstrap values (values ≥ 20) are indicated next to the corresponding branches. The tree specific topology was tested by Maximum-Parsimony analysis. GNAT candidates with a putative organellar localization (TargetP1.1) were highlighted with a green background and named as GNAT1 to GNAT10 in relation to their position in the phylogenetic tree. Squares, triangles and circles describe the specific acetylation activity, which was reported in literature. The metabolic activity of GNAT2 corresponds to serotonin acetyltransferase (B) Schematic overview of organellar GNATs' secondary structure organization (including AtNAA and EcRiml for comparison). Secondary structural elements of the GNAT candidates were determined using Jpred tools in combination with structure homology-models (Swiss-model) and are displayed in red (α-helixes), green (β-strands) and orange (supplementary secondary elements). All candidates were predicted with a mitochondrial or a chloroplastic transit peptide (cTP) using TargetP. The mature form of these candidates is released after the excision of this cTP. Positions of main and secondary Acyl-CoA binding domain (Ac-CoA BD) are shown. C, D, A and B design the four conserved motifs comprising what is referred as the N-acetyltransferase domain