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

J. Biol. Chem., Vol. 280, Issue 8, 6554-6560, February 25, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/8/6554    most recent M412928200v1 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 Google Scholar Google Scholar Articles by Shostak, Y. Articles by Yamamoto, K. R. Search for Related Content PubMed PubMed Citation Articles by Shostak, Y. Articles by Yamamoto, K. R. Social Bookmarking                      What's this?

Overlapping but Separable Determinants of DNA Binding and Nuclear Localization Map to the C-terminal End of the Caenorhabditis elegans DAF-12 DNA Binding Domain^*

Yuriy Shostak and Keith R. Yamamoto

From the Program in Biochemistry and Molecular Biology and Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California 94143-2280

Proteins are commonly viewed as modular assemblies of functional domains. We analyzed a loss-of-function mutation in the Caenorhabditis elegans intracellular receptor DAF-12, a conservative substitution of an arginine to a lysine at position 197 (R197K). Arg¹⁹⁷ resides in region similar to a nuclear localization signal, just downstream of the receptor minimal zinc finger DNA binding domain (DBD) core. We found that the R197K, but not mutations of neighboring arginine or lysine residues, dramatically reduced DAF-12 transcriptional regulatory activity in a yeast reporter assay. This reduction in regulatory activity correlated with greatly decreased DNA binding affinity in vitro, suggesting a role for the DAF-12 DBD C-terminal region (dbdC), and specifically for Arg¹⁹⁷, in DNA binding. Remarkably, three basic residues immediately contiguous with Arg¹⁹⁷ played little role in DNA binding and rather affected nuclear localization; in contrast, Arg¹⁹⁷ itself was dispensable for nuclear localization. Thus, DAF-12 dbdC harbors overlapping but separable determinants of DNA binding and nuclear localization in a single small region.

Received for publication, November 16, 2004

^* This work was supported by National Institutes of Health and National Science Foundation (NSF) grants. 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.

Supported by an NSF graduate research fellowship.

To whom correspondence should be addressed. Tel.: 415-476-3128; Fax: 415-476-6129; E-mail: yamamoto{at}cgl.ucsf.edu.

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