title: COL11A2 collagen gene transcription is differentially regulated by EWS/ERG sarcoma fusion protein and wild-type ERG.
abstract: A specific t(21;22) chromosomal translocation creates the chimeric EWS/ERG gene in some cases of Ewing's sarcoma. In the resultant EWS/ERG fusion protein, the N-terminal part of the ETS family protein ERG is replaced by the N terminus of the RNA-binding protein EWS. We found that both the EWS/ERG and COL11A2 genes are expressed in the Ewing's sarcoma cell line, CADO-ES1. To investigate a potential role for EWS/ERG in COL11A2 gene expression, we characterized the COL11A2 promoter and tested the ability of wild-type ERG and EWS/ERG sarcoma fusion protein to transactivate COL11A2 promoter using a luciferase assay. We found that expression of EWS/ERG, but not wild-type ERG, transactivated the COL11A2 promoter and that this transactivation required not only the N-terminal region of EWS but also an intact DNA-binding domain from ERG. Electrophoretic mobility shift assay using COL11A2 promoter sequence showed involvement of EWS/ERG in the formation of DNA-protein complexes, and chromatin immunoprecipitation assay revealed direct interaction between COL11A2 promoter and EWS/ERG fusion protein in vivo. EWS/ERG, but not wild-type ERG, bound to RNA polymerase II. Treatment of cells with the histone deacetylase inhibitor trichostatin A enabled ERG to transactivate the COL11A2 promoter, therefore abolishing the differential effects of EWS/ERG and ERG. Taken together, these findings indicate that the COL11A2 gene is regulated both by potential ERG association with a histone deacetylase complex and by direct EWS/ERG recruitment of RNA polymerase II.
authors.affiliation: Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington 98195-6500, USA.
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Matsui
authors.forename: Howard A
authors.initials: HA
authors.lastname: Chansky
authors.forename: Fariba
authors.initials: F
authors.lastname: Barahmand-Pour
authors.forename: Anna
authors.initials: A
authors.lastname: Zielinska-Kwiatkowska
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Tsumaki
authors.forename: Akira
authors.initials: A
authors.lastname: Myoui
authors.forename: Hideki
authors.initials: H
authors.lastname: Yoshikawa
authors.forename: Liu
authors.initials: L
authors.lastname: Yang
authors.forename: David R
authors.initials: DR
authors.lastname: Eyre
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003094:Collagen / Q000235:genetics*; D017931:DNA Primers; D004268:DNA-Binding Proteins; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation / Q000502:physiology; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D051379:Mice; D015513:Oncogene Proteins / Q000502:physiology*; D015514:Oncogene Proteins, Fusion / Q000502:physiology*; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*; D000071230:Transcriptional Regulator ERG
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; C072312:ERG protein, human; C450460:EWS-ERG fusion protein, human; D015513:Oncogene Proteins; D015514:Oncogene Proteins, Fusion; D015534:Trans-Activators; D014157:Transcription Factors; D000071230:Transcriptional Regulator ERG; D003094:Collagen; D012319:RNA Polymerase II; D006655:Histone Deacetylases