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

J. Biol. Chem., Vol. 281, Issue 46, 35531-35543, November 17, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/46/35531    most recent M606090200v1 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 Johnson, A. Articles by O'Donnell, M. Search for Related Content PubMed PubMed Citation Articles by Johnson, A. Articles by O'Donnell, M. Related Collections Papers Of The Week Social Bookmarking                      What's this?

The Replication Factor C Clamp Loader Requires Arginine Finger Sensors to Drive DNA Binding and Proliferating Cell Nuclear Antigen Loading^*

Aaron Johnson, Nina Y. Yao, Gregory D. Bowman, John Kuriyan^¶||, and Mike O'Donnell^**1

From the ^**Laboratory of DNA Replication, Howard Hughes Medical Institute and Rockefeller University, New York, New York 10021, the ^¶Howard Hughes Medical Institute, Department of Molecular and Cell Biology and Department of Chemistry, University of California, Berkeley, California 94720, the ^||Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, and the Department of Biophysics, Johns Hopkins University, Baltimore, Maryland 21218

Replication factor C (RFC) is an AAA+ heteropentamer that couples the energy of ATP binding and hydrolysis to the loading of the DNA polymerase processivity clamp, proliferating cell nuclear antigen (PCNA), onto DNA. RFC consists of five subunits in a spiral arrangement (RFC-A, -B, -C, -D, and -E, corresponding to subunits RFC1, RFC4, RFC3, RFC2, and RFC5, respectively). The RFC subunits are AAA+ family proteins and the complex contains four ATP sites (sites A, B, C, and D) located at subunit interfaces. In each ATP site, an arginine residue from one subunit is located near the -phosphate of ATP bound in the adjacent subunit. These arginines act as "arginine fingers" that can potentially perform two functions: sensing that ATP is bound and catalyzing ATP hydrolysis. In this study, the arginine fingers in RFC were mutated to examine the steps in the PCNA loading mechanism that occur after RFC binds ATP. This report finds that the ATP sites of RFC function in distinct steps during loading of PCNA onto DNA. ATP binding to RFC powers recruitment and opening of PCNA and activates a -phosphate sensor in ATP site C that promotes DNA association. ATP hydrolysis in site D is uniquely stimulated by PCNA, and we propose that this event is coupled to PCNA closure around DNA, which starts an ordered hydrolysis around the ring. PCNA closure severs contact to RFC subunits D and E (RFC2 and RFC5), and the -phosphate sensor of ATP site C is switched off, resulting in low affinity of RFC for DNA and ejection of RFC from the site of PCNA loading.

Received for publication, June 26, 2006 , and in revised form, September 14, 2006.

^* This work was supported by National Institutes of Health Grant GM38839 (to M. O. D.), an institutional training grant (to A. J.), the Ruth L. Kirschstein National Research Service Award through the National Institute of General Medical Sciences (to G. D. B.), and by National Institutes of Health Grant GM45547 (to J. K.). 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.

This article was selected as a Paper of the Week.

¹ To whom correspondence should be addressed: 1230 York Ave., Box 228, New York, NY 10021-6399. Tel.: 212-327-7255; Fax: 212-327-7253; E-mail: odonnel{at}mail.rockefeller.edu.

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

This article has been cited by other articles:

				H. Ren, S.-X. Dou, P. Rigolet, Y. Yang, P.-Y. Wang, M. Amor-Gueret, and X. G. Xi The arginine finger of the Bloom syndrome protein: its structural organization and its role in energy coupling Nucleic Acids Res., September 25, 2007; 35(18): 6029 - 6041. [Abstract] [Full Text] [PDF]

				S. G. Anderson, C. R. Williams, M. O'Donnell, and L. B. Bloom A Function for the {psi} Subunit in Loading the Escherichia coli DNA Polymerase Sliding Clamp J. Biol. Chem., March 9, 2007; 282(10): 7035 - 7045. [Abstract] [Full Text] [PDF]