title: Expression of Non-acetylatable H2A.Z in Myoblast Cells Blocks Myoblast Differentiation through Disruption of MyoD Expression.
abstract: H2A.Z is a histone H2A variant that is essential for viability in Tetrahymena and Drosophila and also during embryonic development of mice. Although implicated in diverse cellular processes, including transcriptional regulation, chromosome segregation, and heterochromatin formation, its essential function in cells remains unknown. Cellular differentiation is part of the developmental process of multicellular organisms. To elucidate the roles of H2A.Z and H2A.Z acetylation in cellular differentiation, we examined the effects of expressing wild type (WT) or a non-acetylatable form of H2A.Z in the growth and differentiation of the myoblast C2C12 cell line. Ectopic expression of wild type or mutant H2A.Z resulted in distinct phenotypes in the differentiation of the C2C12 cells and the formation of myotubes. Most strikingly, expression of the H2A.Z non-acetylatable mutant (H2A.Z-Ac-mut) resulted in a complete block of myoblast differentiation. We determined that this phenotype is caused by a loss of MyoD expression in the Ac-mut-expressing cells prior to and after induction of differentiation. Moreover, chromatin accessibility assays showed that the promoter region of MyoD is less accessible in the differentiation-defective cells. Altogether, these new findings show that expression of the Ac-mut form of H2A.Z resulted in a dominant phenotype that blocked differentiation due to chromatin changes at the MyoD promoter.
authors.affiliation: From the Department of Medical Biophysics, University of Toronto, Ontario M5G 1L7, Canada and.
authors.forename: Cindy
authors.initials: C
authors.lastname: Law
authors.affiliation: From the Department of Medical Biophysics, University of Toronto, Ontario M5G 1L7, Canada and the Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada pmcheung@yorku.ca.
authors.forename: Peter
authors.initials: P
authors.lastname: Cheung
mesh_terms: D000107:Acetylation; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D005455:Fluorescent Antibody Technique; D006657:Histones / Q000378:metabolism*; D051379:Mice; D017570:MyoD Protein / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology* / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D006657:Histones; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D009707:Nucleosomes; D012333:RNA, Messenger
keywords: H2A.Z; chromatin; gene regulation; histone modification; histone variant; myogenesis; transcription
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