title: SMARCA4 regulates gene expression and higher-order chromatin structure in proliferating mammary epithelial cells.
abstract: The packaging of DNA into chromatin plays an important role in transcriptional regulation and nuclear processes. Brahma-related gene-1 SMARCA4 (also known as BRG1), the essential ATPase subunit of the mammalian SWI/SNF chromatin remodeling complex, uses the energy from ATP hydrolysis to disrupt nucleosomes at target regions. Although the transcriptional role of SMARCA4 at gene promoters is well-studied, less is known about its role in higher-order genome organization. SMARCA4 knockdown in human mammary epithelial MCF-10A cells resulted in 176 up-regulated genes, including many related to lipid and calcium metabolism, and 1292 down-regulated genes, some of which encode extracellular matrix (ECM) components that can exert mechanical forces and affect nuclear structure. ChIP-seq analysis of SMARCA4 localization and SMARCA4-bound super-enhancers demonstrated extensive binding at intergenic regions. Furthermore, Hi-C analysis showed extensive SMARCA4-mediated alterations in higher-order genome organization at multiple resolutions. First, SMARCA4 knockdown resulted in clustering of intra- and inter-subtelomeric regions, demonstrating a novel role for SMARCA4 in telomere organization. SMARCA4 binding was enriched at topologically associating domain (TAD) boundaries, and SMARCA4 knockdown resulted in weakening of TAD boundary strength. Taken together, these findings provide a dynamic view of SMARCA4-dependent changes in higher-order chromatin organization and gene expression, identifying SMARCA4 as a novel component of chromatin organization.
authors.affiliation: Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA;
authors.forename: A Rasim
authors.initials: AR
authors.lastname: Barutcu
authors.affiliation: Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA;
authors.forename: Bryan R
authors.initials: BR
authors.lastname: Lajoie
authors.affiliation: Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont 05405, USA;
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Fritz
authors.affiliation: Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee 37996, USA;
authors.forename: Rachel P
authors.initials: RP
authors.lastname: McCord
authors.affiliation: Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA;
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Nickerson
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA;
authors.forename: Andre J
authors.initials: AJ
authors.lastname: van Wijnen
authors.affiliation: Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont 05405, USA;
authors.forename: Jane B
authors.initials: JB
authors.lastname: Lian
authors.affiliation: Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont 05405, USA;
authors.forename: Janet L
authors.initials: JL
authors.lastname: Stein
authors.affiliation: Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA; Howard Hughes Medical Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors.forename: Job
authors.initials: J
authors.lastname: Dekker
authors.affiliation: Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont 05405, USA;
authors.forename: Gary S
authors.initials: GS
authors.lastname: Stein
authors.affiliation: Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA;
authors.forename: Anthony N
authors.initials: AN
authors.lastname: Imbalzano
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics*; D002843:Chromatin / Q000235:genetics*; D042002:Chromatin Assembly and Disassembly; D004265:DNA Helicases / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics*; D009707:Nucleosomes / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D002843:Chromatin; D009687:Nuclear Proteins; D009707:Nucleosomes; D014157:Transcription Factors; C084108:SMARCA4 protein, human; D004265:DNA Helicases