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

J. Biol. Chem., Vol. 279, Issue 14, 13616-13623, April 2, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/14/13616    most recent M313202200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Clark, L. B. Articles by Denis, C. L. Search for Related Content PubMed PubMed Citation Articles by Clark, L. B. Articles by Denis, C. L. Social Bookmarking                      What's this?

Systematic Mutagenesis of the Leucine-rich Repeat (LRR) Domain of CCR4 Reveals Specific Sites for Binding to CAF1 and a Separate Critical Role for the LRR in CCR4 Deadenylase Activity^*

Lisa B. Clark, Palaniswamy Viswanathan, Gloria Quigley, Yueh-Chin Chiang, Jeffrey S. McMahon, Gang Yao, Junji Chen, Andreas Nelsbach, and Clyde L. Denis

From the Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham, New Hampshire 03824

CCR4, a poly(A) deadenylase of the exonuclease III family, is a component of the multiprotein CCR4-NOT complex of Saccharomyces cerevisiae that is involved in mRNA degradation. CCR4, unlike all other exonuclease III family members, contains a leucine-rich repeat (LRR) motif through which it makes contact to CAF1 and other factors. The LRR residues important in contacting CAF1 were identified by constructing 29 CCR4 mutations encompassing a majority (47 of 81) of residues interstitial to the conserved structural residues. Two-hybrid and immunoprecipitation data revealed that physical contact between CAF1 and the LRR is blocked by mutation of just two -helix/-helix strand loop residues linking the first and second repeats. In contrast, CAF16, a potential ligand of CCR4, was abrogated in its binding to the LRR by mutations in the N terminus of the second -strand. The LRR domain was also found to contact the deadenylase domain of CCR4, and deletion of the LRR region completely inhibited CCR4 enzymatic activity. Mutations throughout the -sheet surface of the LRR, including those that did not specifically interfere with contacts to CAF1 or CAF16, significantly reduced CCR4 deadenylase activity. These results indicate that the CCR4-LRR, in addition to binding to CAF1, plays an essential role in the CCR4 deadenylation of mRNA.

Received for publication, December 3, 2003

^* This work was supported by National Institutes of Health, Division of General Medical Sciences, Grants 1 F32 GM20235-01 (to L. B. C.) and GM41215 and Hatch 291 (to C. L. D.). This is Publication 2118 from the New Hampshire Agriculture Experimental Station. 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.

To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, Rudman Hall, University of New Hampshire, 46 College Rd., Durham, NH 03824. Tel.: 603-862-2438; Fax: 603-862-4013; E-mail address: cldenis{at}cisunix.unh.edu.

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

This article has been cited by other articles:

				A. Schwede, L. Ellis, J. Luther, M. Carrington, G. Stoecklin, and C. Clayton A role for Caf1 in mRNA deadenylation and decay in trypanosomes and human cells Nucleic Acids Res., June 1, 2008; 36(10): 3374 - 3388. [Abstract] [Full Text] [PDF]

				C. Gelling, I. W. Dawes, N. Richhardt, R. Lill, and U. Muhlenhoff Mitochondrial Iba57p Is Required for Fe/S Cluster Formation on Aconitase and Activation of Radical SAM Enzymes Mol. Cell. Biol., March 1, 2008; 28(5): 1851 - 1861. [Abstract] [Full Text] [PDF]

				F. Abe and H. Minegishi Global Screening of Genes Essential for Growth in High-Pressure and Cold Environments: Searching for Basic Adaptive Strategies Using a Yeast Deletion Library Genetics, February 1, 2008; 178(2): 851 - 872. [Abstract] [Full Text] [PDF]

				T. Ohn, Y.-C. Chiang, D. J. Lee, G. Yao, C. Zhang, and C. L. Denis CAF1 plays an important role in mRNA deadenylation separate from its contact to CCR4 Nucleic Acids Res., May 14, 2007; 35(9): 3002 - 3015. [Abstract] [Full Text] [PDF]

				J. Z. Morris, A. Hong, M. A. Lilly, and R. Lehmann twin, a CCR4 homolog, regulates cyclin poly(A) tail length to permit Drosophila oogenesis Development, March 15, 2005; 132(6): 1165 - 1174. [Abstract] [Full Text] [PDF]

				P. Viswanathan, T. Ohn, Y.-C. Chiang, J. Chen, and C. L. Denis Mouse CAF1 Can Function As a Processive Deadenylase/3'-5'-Exonuclease in Vitro but in Yeast the Deadenylase Function of CAF1 Is Not Required for mRNA Poly(A) Removal J. Biol. Chem., June 4, 2004; 279(23): 23988 - 23995. [Abstract] [Full Text] [PDF]