title: DUX4 recruits p300/CBP through its C-terminus and induces global H3K27 acetylation changes.
abstract: Ectopic expression of the double homeodomain transcription factor DUX4 causes facioscapulohumeral muscular dystrophy (FSHD). Mechanisms of action of DUX4 are currently unknown. Using immortalized human myoblasts with a titratable DUX4 transgene, we identify by mass spectrometry an interaction between the DUX4 C-terminus and the histone acetyltransferases p300/CBP. Chromatin immunoprecipitation shows that DUX4 recruits p300 to its target gene, ZSCAN4, displaces histone H3 from the center of its binding site, and induces H3K27Ac in its vicinity, but C-terminal deleted DUX4 does not. We show that a DUX4 minigene, bearing only the homeodomains and C-terminus, is transcriptionally functional and cytotoxic, and that overexpression of a nuclear targeted C-terminus impairs the ability of WT DUX4 to interact with p300 and to regulate target genes. Genomic profiling of DUX4, histone H3, and H3 modifications reveals that DUX4 binds two classes of loci: DNase accessible H3K27Ac-rich chromatin and inaccessible H3K27Ac-depleted MaLR-enriched chromatin. At this latter class, it acts as a pioneer factor, recruiting H3K27 acetyltransferase activity and opening the locus for transcription. In concert with local increased H3K27Ac, the strong H3K27Ac peaks at distant sites are significantly depleted of H3K27Ac, thus DUX4 uses its C-terminus to induce a global reorganization of H3K27 acetylation.
authors.affiliation: Lillehei Heart Institute, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Research Center, Dongnam Institute of Radiological & Medical Sciences (DIRAMS), Busan, South Korea.
authors.forename: Si Ho
authors.initials: SH
authors.lastname: Choi
authors.affiliation: Department of Genetics, Cell Biology and Development, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA.
authors.forename: Micah D
authors.identifier: 0000-0002-9873-1930
authors.initials: MD
authors.lastname: Gearhart
authors.affiliation: Lillehei Heart Institute, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA.
authors.forename: Ziyou
authors.initials: Z
authors.lastname: Cui
authors.affiliation: Lillehei Heart Institute, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA.
authors.forename: Darko
authors.initials: D
authors.lastname: Bosnakovski
authors.affiliation: Research Center, Dongnam Institute of Radiological & Medical Sciences (DIRAMS), Busan, South Korea.
authors.forename: Minjee
authors.initials: M
authors.lastname: Kim
authors.affiliation: Lillehei Heart Institute, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA.
authors.forename: Natalie
authors.initials: N
authors.lastname: Schennum
authors.affiliation: Lillehei Heart Institute, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, MN 55455, USA kyba@umn.edu.
authors.forename: Michael
authors.initials: M
authors.lastname: Kyba
mesh_terms: D000107:Acetylation; D001665:Binding Sites; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic; D057890:Epigenomics / Q000379:methods; D015870:Gene Expression; D017930:Genes, Reporter; D059014:High-Throughput Nucleotide Sequencing; D006657:Histones / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D032446:Myoblasts / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D014158:Transcription, Genetic; D050880:p300-CBP Transcription Factors / Q000378:metabolism*
chemical_list: C121257:DUX4L1 protein, human; D006657:Histones; D018398:Homeodomain Proteins; D050880:p300-CBP Transcription Factors