title: ARID3B Directly Regulates Ovarian Cancer Promoting Genes.
abstract: The DNA-binding protein AT-Rich Interactive Domain 3B (ARID3B) is elevated in ovarian cancer and increases tumor growth in a xenograft model of ovarian cancer. However, relatively little is known about ARID3B's function. In this study we perform the first genome wide screen for ARID3B direct target genes and ARID3B regulated pathways. We identified and confirmed numerous ARID3B target genes by chromatin immunoprecipitation (ChIP) followed by microarray and quantitative RT-PCR. Using motif-finding algorithms, we characterized a binding site for ARID3B, which is similar to the previously known site for the ARID3B paralogue ARID3A. Functionality of this predicted site was demonstrated by ChIP analysis. We next demonstrated that ARID3B induces expression of its targets in ovarian cancer cell lines. We validated that ARID3B binds to an epidermal growth factor receptor (EGFR) enhancer and increases mRNA expression. ARID3B also binds to the promoter of Wnt5A and its receptor FZD5. FZD5 is highly expressed in ovarian cancer cell lines, and is upregulated by exogenous ARID3B. Both ARID3B and FZD5 expression increase adhesion to extracellular matrix (ECM) components including collagen IV, fibronectin and vitronectin. ARID3B-increased adhesion to collagens II and IV require FZD5. This study directly demonstrates that ARID3B binds target genes in a sequence-specific manner, resulting in increased gene expression. Furthermore, our data indicate that ARID3B regulation of direct target genes in the Wnt pathway promotes adhesion of ovarian cancer cells.
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine-South Bend, South Bend, Indiana, United States of America; Harper Cancer Research Institute, South Bend, Indiana, United States of America.
authors.forename: Alexander
authors.initials: A
authors.lastname: Bobbs
authors.affiliation: Harper Cancer Research Institute, South Bend, Indiana, United States of America; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, United States of America.
authors.forename: Katrina
authors.initials: K
authors.lastname: Gellerman
authors.affiliation: Harper Cancer Research Institute, South Bend, Indiana, United States of America; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, United States of America.
authors.forename: William Morgan
authors.initials: WM
authors.lastname: Hallas
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine-South Bend, South Bend, Indiana, United States of America; Harper Cancer Research Institute, South Bend, Indiana, United States of America.
authors.forename: Stancy
authors.initials: S
authors.lastname: Joseph
authors.affiliation: Harper Cancer Research Institute, South Bend, Indiana, United States of America; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, United States of America.
authors.forename: Chao
authors.initials: C
authors.lastname: Yang
authors.affiliation: Harper Cancer Research Institute, South Bend, Indiana, United States of America; Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, United States of America.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Kurkewich
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine-South Bend, South Bend, Indiana, United States of America; Harper Cancer Research Institute, South Bend, Indiana, United States of America; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, United States of America; Indiana University Melvin and Bren Simon Cancer Center, Indianapolis, Indiana, United States of America.
authors.forename: Karen D
authors.initials: KD
authors.lastname: Cowden Dahl
mesh_terms: D000465:Algorithms; D000818:Animals; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D051157:Frizzled Receptors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019096:Vitronectin / Q000235:genetics / Q000378:metabolism
chemical_list: C120893:ARID3B protein, human; D004268:DNA-Binding Proteins; C498589:FZD5 protein, human; D005353:Fibronectins; D051157:Frizzled Receptors; D012333:RNA, Messenger; D019096:Vitronectin