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Methyl peptide sequence: IVIFASNR(me2)GNCVIR(me2)GTEDITS Non-methyl peptide sequence: IVIFASNRGNCVIRGTEDITS. i The specificity of antibodies against methylated Pontin at R333/R339 residues was assessed by dot blot analysis. Conserved arginine residues (R333/R339) are highlighted with red. h Sequence alignment of human Pontin with selected homologs. g Ribbon diagram of Pontin (PDB 2C9O) showing its structure with R333 and R339 residues. f In vitro methylation assay of Pontin WT, R333K/R339K mutant, or R333A/R339A mutant proteins was performed with CARM1 proteins. Immunoblot analysis with anti-Rme2a antibody was performed after GST pulldown. e In vitro methylation assay using purified GST-Pontin WT, R333A, R339A, or R333/339A mutant with HA-CARM1. Immunoblotting with anti-asymmetric dimethyl arginine (Rme2a) antibody was performed after Ni-NTA pulldown. d In vitro methylation assay using purified His-Pontin was performed with either HA-CARM1 WT or enzymatic activity-deficient R169A mutant. c GST-pulldown assay for binding of CARM1 to Pontin and Reptin. b The binding of Pontin to CARM1 was detected from the eluate of affinity chromatography by immunoblot analysis. Mass spectrometric analysis identified Pontin as a CARM1-interacting protein under glucose starvation. Together, our findings provide a signaling axis of CARM1-Pontin-FOXO3a and further expand the role of CARM1 in nuclear regulation of autophagy.Ī CARM1-interacting proteins were purified from HEK293T cells stably expressing Flag-CARM1 by co-immunoprecipitation with Flag-M2 agarose. Surprisingly, CARM1-Pontin-FOXO3a signaling axis can work in the distal regions and activate autophagy genes through enhancer activation. Genome-wide analyses and biochemical studies reveal that methylated Pontin functions as a platform for recruiting Tip60 histone acetyltransferase with increased H4 acetylation and subsequent activation of autophagy genes regulated by FOXO3a. Here, we identify that coactivator-associated arginine methyltransferase 1 (CARM1) methylates Pontin chromatin-remodeling factor under glucose starvation, and methylated Pontin binds Forkhead Box O 3a (FOXO3a). Recently, transcriptional and epigenetic regulations of autophagy have emerged as essential mechanisms for maintaining homeostasis. is a catabolic process through which cytoplasmic components are degraded and recycled in response to various stresses including starvation.
#Essential anatomy 3 activation code code
7 Creative Research Initiatives Center for Epigenetic Code and Diseases, Department of Biological Sciences, Seoul National University, Seoul, 08826, South Korea.6 Novo Nordisk Foundation Center for Stem Cell Biology, DanStem, Faculty of Health and Medical Sciences, University of Copenhagen, 2200, Copenhagen N, Denmark.5 Biotech Research and Innovation Centre (BRIC), University of Copenhagen, 2200, Copenhagen N, Denmark.4 Department of Molecular Cell Biology, School of Medicine, Sungkyunkwan University, Suwon, 16419, South Korea.3 Department of Anatomy, College of Medicine, The Catholic University of Korea, Seoul, 06591, South Korea.2 Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94720, USA.1 Creative Research Initiatives Center for Epigenetic Code and Diseases, Department of Biological Sciences, Seoul National University, Seoul, 08826, South Korea.