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Journal of Biochemistry Advance Access originally published online on November 26, 2007
Journal of Biochemistry 2008 143(3):395-406; doi:10.1093/jb/mvm231
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© 2007 The Japanese Biochemical Society.

Target Specificities of Estrogen Receptor-Related Receptors: Analysis of Binding Sequences and Identification of Rb1-Inducible Coiled-Coil 1 (Rb1cc1) as a Target Gene

Mst. Hasina Akter1, Tokuhiro Chano2, Hidetoshi Okabe2, Tomohiro Yamaguchi1, Fumiko Hirose1 and Takashi Osumi1,*

1Graduate School of Life Science, University of Hyogo, Kamigori, Hyogo 678-1297; and 2Department of Clinical Laboratory Medicine, Shiga University of Medical Science, Seta Tsukinowa-cho, Otsu, Shiga 520-2192, Japan

*To whom correspondence should be addressed: Tel: +81-791-58-0192, Fax: +81-791-58-0193, E-mail: osumi{at}sci.u-hyogo.ac.jp

Received October 1, 2007; Accepted November 21, 2007


  Abstract

Estrogen receptor-related receptors (ERRs) are orphan members of the nuclear receptor superfamily. A single AGGTCA sequence element preceded by three conserved nucleotides has been identified as a specific recognition motif of ERRs. Here we performed systematic analyses of target sequences on all three ERR subtypes, {alpha}, β and {gamma}. In electrophoretic gel-mobility shift assay and transcriptional reporter assays, they exhibited similar patterns of recognition specificities, showing extremely broad ranges of target sequences. We searched a mouse promoter database for a gene carrying possible ERR-binding sequences. The Rb-1 inducible coiled-coil 1 (Rb1cc1) gene was found to contain two putative ERR binding elements, named response element (RE)-1 and RE-2, in the promoter region. In gene reporter assays, RE-2, but not RE-1, functioned as an effective cis-regulatory element for transactivation by ERR{alpha} in the presence of a coactivator, peroxisome proliferator-activated receptor {gamma} coactivator-1{alpha}. Mutational analyses suggested that RE-2 is recognized by ERR{alpha} partly as a monovalent element, but also as a direct repeat motif separated by four spacer nucleotides. In vivo binding of ERR{alpha} to the Rb1cc1 promoter region was confirmed by the chromatin immunoprecipitation assay. Thus, Rb1cc1 is a target gene of ERR{alpha}, driven by a novel type of recognition sequence.

Key Words: estrogen receptor-related receptor, nuclear hormone receptor, Rb1-inducible coiled-coil 1, response element, target gene, transcriptional regulation

Abbreviations: ChIP, chromatin immunoprecipitation; DR, direct repeat; EMSA, electrophoretic-gel mobility-shift assay; ER, estrogen receptor; ERR, estrogen receptor-related receptor; IR, inverted repeat; PGC-1{alpha}, peroxisome proliferator-activated receptor {gamma} coactivator-1 {alpha}; Rb1cc1, Rb1-inducible coiled-coil 1; RE, response element


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