HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 36, 31818-31829, September 9, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/36/31818    most recent M501924200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gu, P. Articles by Cooney, A. J. Search for Related Content PubMed PubMed Citation Articles by Gu, P. Articles by Cooney, A. J. Social Bookmarking                      What's this?

Evolutionary Trace-based Peptides Identify a Novel Asymmetric Interaction That Mediates Oligomerization in Nuclear Receptors^*

Peili Gu, Daniel H. Morgan¶||, Minawar Sattar, Xueping Xu, Ryan Wagner, Michele Raviscioni¶**, Olivier Lichtarge¶||**, and Austin J. Cooney

From the Department of Molecular and Cellular Biology, ^¶Department of Molecular and Human Genetics, the ^||Graduate Program in Structural and Computational Biology and Molecular Biophysics, and the ^**W. M. Keck Center for Computational and Structural Biology, Baylor College of Medicine, Houston, Texas 77030

Germ cell nuclear factor (GCNF) is an orphan nuclear receptor that plays important roles in development and reproduction, by repressing the expression of essential genes such as Oct4, GDF9, and BMP15, through binding to DR0 elements. Surprisingly, whereas recombinant GCNF binds to DR0 sequences as a homodimer, endogenous GCNF does not exist as a homodimer but rather as part of a large complex termed the transiently retinoid-induced factor (TRIF). Here, we use evolutionary trace (ET) analysis to design mutations and peptides that probe the molecular basis for the formation of this unusual complex. We find that GCNF homodimerization and TRIF complex formation are DNA-dependent, and ET suggests that dimerization involves key functional sites on both helix 3 and helix 11, which are located on opposing surfaces of the ligand binding domain. Targeted mutations in either helix of GCNF disrupt the formation of both the homodimer and the endogenous TRIF complex. Moreover, peptide mimetics of both of these ET-determined sites inhibit dimerization and TRIF complex formation. This suggests that a novel helix 3-helix 11 heterotypic interaction mediates GCNF interaction and would facilitate oligomerization. Indeed, it was determined that the endogenous TRIF complex is composed of a GCNF oligomer. These findings shed light on an evolutionarily selected mechanism that reveals the unusual DNA-binding, dimerization, and oligomerization properties of GCNF.

Received for publication, February 22, 2005 , and in revised form, June 22, 2005.

^* This work was supported by National Institutes of Health Nuclear Receptor Signaling Atlas orphan receptor program Grant U19DK6234-01 and in part by National Science Foundation Grant DBI-0318415, National Institutes of Health Grant R01 GM066099, and March of Dimes Grant 1-FY03-93 (to O. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

These authors contributed equally to this work.

Supported by a training fellowship from the W. M. Keck Center for Computational and Structural Biology.

To whom correspondence should be addressed: Dept. of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Tel.: 713-798-6250; Fax: 713-790-1275; E-mail: acooney{at}bcm.tmc.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Liu, S. T. Whitten, and V. J. Hilser Functional residues serve a dominant role in mediating the cooperativity of the protein ensemble PNAS, March 13, 2007; 104(11): 4347 - 4352. [Abstract] [Full Text] [PDF]

				P. Gu, D. Le Menuet, A. C.-K. Chung, and A. J. Cooney Differential Recruitment of Methylated CpG Binding Domains by the Orphan Receptor GCNF Initiates the Repression and Silencing of Oct4 Expression Mol. Cell. Biol., December 15, 2006; 26(24): 9471 - 9483. [Abstract] [Full Text] [PDF]

				G. Benoit, A. Cooney, V. Giguere, H. Ingraham, M. Lazar, G. Muscat, T. Perlmann, J.-P. Renaud, J. Schwabe, F. Sladek, et al. International Union of Pharmacology. LXVI. Orphan Nuclear Receptors Pharmacol. Rev., December 1, 2006; 58(4): 798 - 836. [Abstract] [Full Text] [PDF]

				M. Hentschke, I. Kurth, U. Borgmeyer, and C. A. Hubner Germ Cell Nuclear Factor Is a Repressor of CRIPTO-1 and CRIPTO-3 J. Biol. Chem., November 3, 2006; 281(44): 33497 - 33504. [Abstract] [Full Text] [PDF]

				D. H. Morgan, D. M. Kristensen, D. Mittelman, and O. Lichtarge ET viewer: an application for predicting and visualizing functional sites in protein structures Bioinformatics, August 15, 2006; 22(16): 2049 - 2050. [Abstract] [Full Text] [PDF]