Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (266K) Freely available
Right arrow Supplementary Material
Right arrow A corrigendum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (4)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Salles-Passador, I.
Right arrow Articles by Fotedar, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Salles-Passador, I.
Right arrow Articles by Fotedar, R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2003, Vol. 31, No. 17 5202-5211
© 2003 Oxford University Press

Phosphorylation of the PCNA binding domain of the large subunit of replication factor C on Thr506 by cyclin-dependent kinases regulates binding to PCNA

Isabelle Salles-Passador1, Anil Munshi2, Dominique Cannella1, Vincent Pennaneach1,2, Stephane Koundrioukoff1, Michel Jaquinod1, Eric Forest1, Vladimir Podust3, Arun Fotedar2 and Rati Fotedar*,1

1 Institut de Biologie Structurale, J.-P. Ebel, 41 rue Jules Horowitz, F-38027 Grenoble Cedex 1, France, 2 Sidney Kimmel Cancer Center, 10835 Altman Road, San Diego, CA 92121, USA and 3 Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235, USA

*To whom correspondence should be addressed. Tel: +33 4 38 78 96 15; Fax: +33 4 38 78 54 94; Email: fotedar{at}ibs.fr
Correspondence may also be addressed to Arun Fotedar. Tel: +1 858 450 5990; Fax: +1 858 450 3251; Email: afotedar{at}skcc.org
Present address:
Stephane Koundrioukoff, Institute of Veterinary Biochemistry and Molecular Biology, University of Zurich Irchel, Zurich, Switzerland

Replication factor C (RF-C) complex binds to DNA primers and loads PCNA onto DNA, thereby increasing the processivity of DNA polymerases. We have previously identified a distinct region, domain B, in the large subunit of human RF-C (RF-Cp145) which binds to PCNA. We show here that the functional interaction of RF-Cp145 with PCNA is regulated by cdk-cyclin kinases. Phosphorylation of either RF-Cp145 as a part of the RF-C complex or RF-Cp145 domain B by cdk-cyclin kinases inhibits their ability to bind PCNA. A cdk-cyclin phosphorylation site, Thr506 in RF-Cp145, identified by mass spectrometry, is also phosphorylated in vivo. A Thr506->Ala RF-Cp145 domain B mutant is a poor in vitro substrate for cdk-cyclin kinase and, consequently, the ability of this mutant to bind PCNA was not suppressed by phosphorylation. By generating an antibody directed against phospho-Thr506 in RF-Cp145, we demonstrate that phosphorylation of endogenous RF-Cp145 at Thr506 is mediated by CDKs since it is abolished by treatment of cells with the cdk-cyclin inhibitor roscovitine. We have thus mapped an in vivo cdk-cyclin phosphorylation site within the PCNA binding domain of RF-Cp145.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.