Advertisement
JBC

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


     


Originally published In Press as doi:10.1074/jbc.M311272200 on December 9, 2003

J. Biol. Chem., Vol. 279, Issue 8, 6354-6363, February 20, 2004
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
279/8/6354    most recent
M311272200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Dou, S.-X.
Right arrow Articles by Xi, X. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dou, S.-X.
Right arrow Articles by Xi, X. G.
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?

The DNA Binding Properties of the Escherichia coli RecQ Helicase*

Shuo-Xing Dou{ddagger}, Peng-Ye Wang{ddagger}, Hou Qiang Xu§, and Xu Guang Xi§

From the {ddagger}Laboratory of Soft Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100080, China and §Laboratoire de Biotechnologies et Pharmacologie Génétique Appliquée CNRS UMR 8113, Ecole Normale Supérieure de Cachan, 61 avenue du Président Wilson, 94235 Cachan cedex, France

The RecQ helicase family is highly conserved from bacteria to men and plays a conserved role in the preservation of genome integrity. Its deficiency in human cells leads to a marked genomic instability that is associated with premature aging and cancer. To determine the thermodynamic parameters for the interaction of Escherichia coli RecQ helicase with DNA, equilibrium binding studies have been performed using the thermodynamic rigorous fluorescence titration technique. Steady-state fluorescence anisotropy measurements of fluorescein-labeled oligonucleotides revealed that RecQ helicase bound to DNA with an apparent binding stoichiometry of 1 protein monomer/10 nucleotides. This stoichiometry was not altered in the presence of AMPPNP (adenosine 5'-({beta},{gamma}-imido) triphosphate) or ADP. Analyses of RecQ helicase interactions with oligonucleotides of different lengths over a wide range of pH, NaCl, and nucleic acid concentrations indicate that the RecQ helicase has a single strong DNA binding site with an association constant at 25 °C of K = 6.7 ± 0.95 x 106 M-1 and a cooperativity parameter of {omega} = 25.5 ± 1.2. Both single-stranded DNA and double-stranded DNA bind competitively to the same site. The intrinsic affinities are salt-dependent, and the formation of DNA-helicase complex is accompanied by a net release of 3–4 ions. Allosteric effects of nucleotide cofactors on RecQ binding to DNA were observed only for single-stranded DNA in the presence of 1.5 mM AMPPNP, whereas both AMPPNP and ADP had no detectable effect on double-stranded DNA binding over a large range of nucleotide cofactor concentrations.


Received for publication, October 14, 2003 , and in revised form, December 7, 2003.

* This work was supported by the CNRS (to X. G. X.), the National Natural Science Foundation of China Grants 60025516 and 10334100, and the Innovation Project of the Chinese Academy of Sciences. 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.

To whom correspondence should be addressed. Tel.: 33-1-47-40-68-92; Fax: 33-1-47-40-76-71; E-mail: xi{at}lbpa.ens-cachan.fr.


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 BiochemHome page
B. Zhang, A.-h. Zhang, L. Chen, and X. G. Xi
Inhibition of DNA Helicase, ATPase and DNA-Binding Activities of E. coli RecQ Helicase by Chemotherapeutic Agents
J. Biochem., June 1, 2008; 143(6): 773 - 779.
[Abstract] [Full Text] [PDF]


Home page
Nucleic Acids ResHome page
Y. Yang, S.-X. Dou, H. Ren, P.-Y. Wang, X.-D. Zhang, M. Qian, B.-Y. Pan, and X. G. Xi
Evidence for a functional dimeric form of the PcrA helicase in DNA unwinding
Nucleic Acids Res., April 1, 2008; 36(6): 1976 - 1989.
[Abstract] [Full Text] [PDF]


Home page
Nucleic Acids ResHome page
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]


Home page
Nucleic Acids ResHome page
R.-B. Guo, P. Rigolet, H. Ren, B. Zhang, X.-D. Zhang, S.-X. Dou, P.-Y. Wang, M. Amor-Gueret, and X. G. Xi
Structural and functional analyses of disease-causing missense mutations in Bloom syndrome protein
Nucleic Acids Res., September 25, 2007; 35(18): 6297 - 6310.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
X.-D. Zhang, S.-X. Dou, P. Xie, J.-S. Hu, P.-Y. Wang, and X. G. Xi
Escherichia coli RecQ Is a Rapid, Efficient, and Monomeric Helicase
J. Biol. Chem., May 5, 2006; 281(18): 12655 - 12663.
[Abstract] [Full Text] [PDF]


Home page
Nucleic Acids ResHome page
R.-b. Guo, P. Rigolet, L. Zargarian, S. Fermandjian, and X. G. Xi
Structural and functional characterizations reveal the importance of a zinc binding domain in Bloom's syndrome helicase
Nucleic Acids Res., June 1, 2005; 33(10): 3109 - 3124.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
J. L. Liu, P. Rigolet, S.-X. Dou, P.-Y. Wang, and X. G. Xi
The Zinc Finger Motif of Escherichia coli RecQ Is Implicated in Both DNA Binding and Protein Folding
J. Biol. Chem., October 8, 2004; 279(41): 42794 - 42802.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
D. Arosio, S. Costantini, Y. Kong, and A. Vindigni
Fluorescence Anisotropy Studies on the Ku-DNA Interaction: ANION AND CATION EFFECTS
J. Biol. Chem., October 8, 2004; 279(41): 42826 - 42835.
[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 © 2004 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement