Advertisement
JBC

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


     

Originally published In Press as doi:10.1074/jbc.M007032200 on October 6, 2000

J. Biol. Chem., Vol. 276, Issue 1, 114-124, January 5, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/1/114    most recent
M007032200v1
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 Skinner, L. M.
Right arrow Articles by Katzman, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Skinner, L. M.
Right arrow Articles by Katzman, M.
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?

Nucleophile Selection for the Endonuclease Activities of Human, Ovine, and Avian Retroviral Integrases*

Lynn M. SkinnerDagger , Malgorzata SudolDagger , Amy L. Harper§, and Michael KatzmanDagger §

From the Dagger  Department of Medicine and § Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Center, Hershey, Pennsylvania 17033

Retroviral integrases catalyze four endonuclease reactions (processing, joining, disintegration, and nonspecific alcoholysis) that differ in specificity for the attacking nucleophile and target DNA sites. To assess how the two substrates of this enzyme affect each other, we performed quantitative analyses, in three retroviral systems, of the two reactions that use a variety of nucleophiles. The integrase proteins of human immuno- deficiency virus type 1, visna virus, and Rous sarcoma virus exhibited distinct preferences for water or other nucleophiles during site-specific processing of viral DNA and during nonspecific alcoholysis of nonviral DNA. Although exogenous alcohols competed with water as the nucleophile for processing, the alcohols stimulated nicking of nonviral DNA. Moreover, different nucleophiles were preferred when the various integrases acted on different DNA targets. In contrast, the nicking patterns were independent of whether integrase was catalyzing hydrolysis or alcoholysis and were not influenced by the particular exogenous alcohol. Thus, although the target DNA influenced the choice of nucleophile, the nucleophile did not affect the choice of target sites. These results indicate that interaction with target DNA is the critical step before catalysis and suggest that integrase does not reach an active conformation until target DNA has bound to the enzyme.

* This work was supported by W. W. Smith Charitable Trust Research Grant A9804, by United States Public Health Services Grant R21-AI47216 from NIAID, the National Institutes of Health, and by the Julius H. Caplan Foundation (in honor of Helen Caplan).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.

To whom correspondence should be addressed: Dept. of Medicine, Division of Infectious Diseases, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Ctr., P. O. Box 850, Mail Code H036, Hershey, PA 17033-0850. Tel.: 717-531-8881; Fax: 717-531-4633; E-mail: mkatzman@psu.edu.

Copyright © 2001 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
 Nucleic Acids ResHome page
T. L. Diamond and F. D. Bushman
Role of metal ions in catalysis by HIV integrase analyzed using a quantitative PCR disintegration assay
Nucleic Acids Res., December 4, 2006; 34(21): 6116 - 6125.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Hizi and H. L. Levin
The Integrase of the Long Terminal Repeat-Retrotransposon Tf1 Has a Chromodomain That Modulates Integrase Activities
J. Biol. Chem., November 25, 2005; 280(47): 39086 - 39094.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
W. M. Konsavage Jr., S. Burkholder, M. Sudol, A. L. Harper, and M. Katzman
A Substitution in Rous Sarcoma Virus Integrase That Separates Its Two Biologically Relevant Enzymatic Activities
J. Virol., April 15, 2005; 79(8): 4691 - 4699.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
A. L. Harper, M. Sudol, and M. Katzman
An Amino Acid in the Central Catalytic Domain of Three Retroviral Integrases That Affects Target Site Selection in Nonviral DNA
J. Virol., March 15, 2003; 77(6): 3838 - 3845.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. K. Bao, A. M. Skalka, and I. Wong
Presteady-state Analysis of Avian Sarcoma Virus Integrase. I. A SPLICING ACTIVITY AND STRUCTURE-FUNCTION IMPLICATIONS FOR COGNATE SITE RECOGNITION
J. Biol. Chem., March 29, 2002; 277(14): 12089 - 12098.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
A. L. Harper, L. M. Skinner, M. Sudol, and M. Katzman
Use of Patient-Derived Human Immunodeficiency Virus Type 1 Integrases To Identify a Protein Residue That Affects Target Site Selection
J. Virol., August 15, 2001; 75(16): 7756 - 7762.
[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 © 2001 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement