/sutils/genom_table.cgi?. We acknowledge the contribution of Karol Romero Merlano for cloning with the rtpD gene and overproduction with the encoded ribitol-5-P 2-dehydrogenase.23. 24.
HHS Public AccessAuthor manuscriptNat Cell Biol. Author manuscript; accessible in PMC 2014 January 01.Published in final edited form as: Nat Cell Biol. 2013 July ; 15(7): 872?82. doi:10.1038/ncb2768.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptProteomic and genomic approaches reveal essential functions of H3K9 methylation and Heterochromatin Protein-1 in reprogramming to pluripotencyRupa Sridharan1,two, Michelle Gonzales-Cope3, , Constantinos Chronis1, , Giancarlo Bonora1, Robin McKee1, Chengyang Huang1, Sanjeet Patel1, David Lopez1, Nilamadhab Mishra4, Matteo Pellegrini1, Michael Carey1, Benjamin A. Garcia3,#, and Kathrin Plath1,#1Universityof California Los Angeles, David Geffen College of Medicine, Division of Biological Chemistry, Jonsson Extensive Cancer Center, Molecular Biology Institute, Bioinformatics Interdepartmental Degree System, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research3EpigeneticsProgram, Division of Biochemistry and Biophysics, Perelman College of Medicine, University of Pennsylvania, 1009C Stellar-Chance Laboratories, 422 Curie Boulevard, Philadelphia, PA, 19104, USA4Sectionon Rheumatology, Division of Internal Medicine, Wake Forest University College of Medicine, Winston-Salem, North CarolinaAbstractReprogramming of somatic cells into iPSCs involves a dramatic reorganization of chromatin.2′-Deoxyadenosine uses To determine posttranslational histone modifications that alter in international abundance throughout this process, we’ve applied a quantitative mass-spectrometry-based strategy.1376340-66-7 Formula We found that iPSCs, in comparison with both the starting fibroblasts in addition to a late reprogramming intermediate (pre-iPSCs), are enriched for histone modifications connected with active chromatin, and depleted for marks of transcriptional elongation and also a subset of repressive modifications such as H3K9me2/me3.PMID:27217159 Dissecting the contribution of H3K9methylation to reprogramming, we show that the H3K9methyltransferases Ehmt1, Ehmt2, and Setdb1 regulate global H3K9me2/me3 levels and that their depletion increases iPSC formation from each fibroblasts and pre-iPSCs. Similarly, inhibition of heterochromatin-protein-1 (Cbx3), a protein identified to recognize H3K9methylation, enhances reprogramming. Genome-wide location analysis revealed that Cbx3 predominantly binds active genes in each pre-iPSCs and pluripotent cells but using a strikingly distinct distribution: in pre-Users may perhaps view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic study, topic generally towards the complete Circumstances of use: http://nature/authors/editorial_policies/license.html#terms # Correspondence: Benjamin Garcia and Kathrin Plath, [email protected], [email protected]. 2Current address: Wisconsin Institute for Discovery, Division of Cell and Regenerative Biology, University of Wisconsin, 330 N. Orchard Street, Area 2118, Madison, WI 53715 These authors contributed equally to this work. Author contributions: R.S., K.P., and B.A.G. planned the project. R.S. and K.P. wrote the manuscript. The following performed experiments, analyzed and interpreted information: R.S., C.C., G.B., R.M., S.P. below K.P.’s supervision, M.G-C. below B.A.G’s supervision, C.H. below M.C.’s supervision, and D.L. beneath M.P.’s supervision. N.M. ge.
