title: Profiling genome-wide chromatin methylation with engineered posttranslation apparatus within living cells.
abstract: Protein methyltransferases (PMTs) have emerged as important epigenetic regulators in myriad biological processes in both normal physiology and disease conditions. However, elucidating PMT-regulated epigenetic processes has been hampered by ambiguous knowledge about in vivo activities of individual PMTs particularly because of their overlapping but nonredundant functions. To address limitations of conventional approaches in mapping chromatin modification of specific PMTs, we have engineered the chromatin-modifying apparatus and formulated a novel technology, termed clickable chromatin enrichment with parallel DNA sequencing (CliEn-seq), to probe genome-wide chromatin modification within living cells. The three-step approach of CliEn-seq involves in vivo synthesis of S-adenosyl-L-methionine (SAM) analogues from cell-permeable methionine analogues by engineered SAM synthetase (methionine adenosyltransferase or MAT), in situ chromatin modification by engineered PMTs, subsequent enrichment and sequencing of the uniquely modified chromatins. Given critical roles of the chromatin-modifying enzymes in epigenetics and structural similarity among many PMTs, we envision that the CliEn-seq technology is generally applicable in deciphering chromatin methylation events of individual PMTs in diverse biological settings.
authors.affiliation: Molecular Pharmacology and Chemistry Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.forename: Kabirul
authors.initials: K
authors.lastname: Islam
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Weihong
authors.initials: W
authors.lastname: Zheng
authors.forename: Haiping
authors.initials: H
authors.lastname: Tang
authors.forename: Nathalie
authors.initials: N
authors.lastname: Lailler
authors.forename: Gil
authors.initials: G
authors.lastname: Blum
authors.forename: Haiteng
authors.initials: H
authors.lastname: Deng
authors.forename: Minkui
authors.initials: M
authors.lastname: Luo
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D057890:Epigenomics; D005818:Genetic Engineering; D052216:Glucagon-Like Peptide 1 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D057809:HEK293 Cells; D006649:Histocompatibility Antigens / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D011499:Protein Processing, Post-Translational; D012436:S-Adenosylmethionine / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D002843:Chromatin; D006649:Histocompatibility Antigens; D012436:S-Adenosylmethionine; D052216:Glucagon-Like Peptide 1; C445405:EHMT2 protein, human; D011495:Histone-Lysine N-Methyltransferase