title: Acetylation of Mammalian ADA3 Is Required for Its Functional Roles in Histone Acetylation and Cell Proliferation.
abstract: Alteration/deficiency in activation 3 (ADA3) is an essential component of specific histone acetyltransferase (HAT) complexes. We have previously shown that ADA3 is required for establishing global histone acetylation patterns and for normal cell cycle progression (S. Mohibi et al., J Biol Chem 287:29442-29456, 2012, http://dx.doi.org/10.1074/jbc.M112.378901). Here, we report that these functional roles of ADA3 require its acetylation. We show that ADA3 acetylation, which is dynamically regulated in a cell cycle-dependent manner, reflects a balance of coordinated actions of its associated HATs, GCN5, PCAF, and p300, and a new partner that we define, the deacetylase SIRT1. We use mass spectrometry and site-directed mutagenesis to identify major sites of ADA3 acetylated by GCN5 and p300. Acetylation-defective mutants are capable of interacting with HATs and other components of HAT complexes but are deficient in their ability to restore ADA3-dependent global or locus-specific histone acetylation marks and cell proliferation in Ada3-deleted murine embryonic fibroblasts (MEFs). Given the key importance of ADA3-containing HAT complexes in the regulation of various biological processes, including the cell cycle, our study presents a novel mechanism to regulate the function of these complexes through dynamic ADA3 acetylation.
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Shakur
authors.initials: S
authors.lastname: Mohibi
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Shashank
authors.initials: S
authors.lastname: Srivastava
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Aditya
authors.initials: A
authors.lastname: Bele
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Sameer
authors.initials: S
authors.lastname: Mirza
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA Department of Biochemistry & Molecular Biology, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA Department of Pathology & Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA Eppley Institute for Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Hamid
authors.initials: H
authors.lastname: Band
authors.affiliation: Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA Eppley Institute for Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, Nebraska, USA vband@unmc.edu.
authors.forename: Vimla
authors.initials: V
authors.lastname: Band
mesh_terms: D000072283:A549 Cells; D000107:Acetylation; D000818:Animals; D001665:Binding Sites; D002453:Cell Cycle; D002460:Cell Line; D049109:Cell Proliferation; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D016297:Mutagenesis, Site-Directed; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D050880:p300-CBP Transcription Factors / Q000378:metabolism*
chemical_list: D006657:Histones; C444103:TADA3 protein, human; C580756:Tada3 protein, mouse; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors