JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M109053200 on November 6, 2001

J. Biol. Chem., Vol. 277, Issue 3, 1719-1727, January 18, 2002
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
277/3/1719    most recent
M109053200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ranalli, T. A.
Right arrow Articles by Bambara, R. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ranalli, T. A.
Right arrow Articles by Bambara, R. A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Mechanism Underlying Replication Protein A Stimulation of DNA Ligase I*

Tamara A. Ranalli, Michael S. DeMottDagger , and Robert A. Bambara§

From the Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642

Replication protein A (RPA) is a heterotrimeric single-stranded DNA-binding protein that participates in multiple DNA transactions that include replication and repair. Base excision repair is a central DNA repair pathway, responsible for the removal of damaged bases. We have shown previously that RPA was able to stimulate long patch base excision repair reconstituted in vitro. Herein we show that human RPA stimulates the activity of the base excision repair component human DNA ligase I by approximately 15-fold. Other analyzed single-stranded binding proteins would not substitute, attesting to the specificity of the stimulation. Conversely, RPA was unable to stimulate the functionally homologous ATP-dependent ligase from T4 bacteriophage. Kinetic analyses suggest that catalysis of ligation is enhanced by RPA, as a 4-fold increase in kcat is observed, whereas Km is not significantly changed. Substrate competition experiments further support the conclusion that RPA does not alter the specificity or rate of substrate binding by DNA ligase I. Additionally, RPA is unable to significantly enhance ligation on substrates containing an unannealed 3'-upstream primer terminus, suggesting that RPA does not stabilize the nick site to enhance ligase recognition. Furthermore when DNA ligase I is pre-bound to the substrate and limited to a single turnover, RPA is still able to stimulate ligation. Overall, the results support a mechanism of stimulation that involves increasing the rate of catalysis of ligation.

* This work was supported by National Institutes of Health Grant GM24441.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Current address: Harvard School of Public Health, Boston, MA 02115.

§ To whom all correspondence should be addressed: Dept. of Biochemistry and Biophysics, Univ. of Rochester Medical Center, 601 Elmwood Ave., Box 712, Rochester, NY 14642. Tel.: 716-275-3269; Fax: 716-271-2683; E-mail: robert_bambara@urmc.rochester.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
J. D. Bartos, L. J. Willmott, S. K. Binz, M. S. Wold, and R. A. Bambara
Catalysis of Strand Annealing by Replication Protein A Derives from Its Strand Melting Properties
J. Biol. Chem., August 1, 2008; 283(31): 21758 - 21768.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. D. Bartos, W. Wang, J. E. Pike, and R. A. Bambara
Mechanisms by Which Bloom Protein Can Disrupt Recombination Intermediates of Okazaki Fragment Maturation
J. Biol. Chem., October 27, 2006; 281(43): 32227 - 32239.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Epidemiol. Biomarkers Prev.Home page
B. A. Sokhansanj and D. M. Wilson III
Estimating the effect of human base excision repair protein variants on the repair of oxidative DNA base damage.
Cancer Epidemiol. Biomarkers Prev., May 1, 2006; 15(5): 1000 - 1008.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Fan, Y. Matsumoto, and D. M. Wilson III
Nucleotide Sequence and DNA Secondary Structure, as Well as Replication Protein A, Modulate the Single-stranded Abasic Endonuclease Activity of APE1
J. Biol. Chem., February 17, 2006; 281(7): 3889 - 3898.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. Wong, M. S. DeMott, and B. Demple
Modulation of the 3'->5'-Exonuclease Activity of Human Apurinic Endonuclease (Ape1) by Its 5'-incised Abasic DNA Product
J. Biol. Chem., September 19, 2003; 278(38): 36242 - 36249.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. A. Ranalli, S. Tom, and R. A. Bambara
AP Endonuclease 1 Coordinates Flap Endonuclease 1 and DNA Ligase I Activity in Long Patch Base Excision Repair
J. Biol. Chem., October 25, 2002; 277(44): 41715 - 41724.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.