Skip Navigation

Nucleic Acids Research 2005 33(15):4865-4873; doi:10.1093/nar/gki779
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (2401K) Freely available
Right arrow Screen PDF (445K) Freely available
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 (5)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Mulder, B. A.
Right arrow Articles by Hardin, S. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mulder, B. A.
Right arrow Articles by Hardin, S. H.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 1 September 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Nucleotide modification at the {gamma}-phosphate leads to the improved fidelity of HIV-1 reverse transcriptase

Brent A. Mulder1,4, Steve Anaya1, Peilin Yu2, Keun Woo Lee1, Anvy Nguyen1,4, Jason Murphy3, Richard Willson1,3,4, James M. Briggs1,4, Xiaolian Gao1,2 and Susan H. Hardin1,4,*

1Department of Biology and Biochemistry, University of Houston Houston TX 77204-5001, USA 2Department of Chemistry, University of Houston Houston TX 77204-5003, USA 3Department of Chemical Engineering, University of Houston Houston, TX 77204-4004, USA 4VisiGen Biotechnologies, Inc. 2575 West Bellfort, Suite 250, Houston, TX 77054, USA

*To whom correspondence should be addressed. Tel: +1 713 743 2686; Fax: +1 713 743 2636; Email: shardin{at}uh.edu

Received May 3, 2005. Revised August 2, 2005. Accepted August 2, 2005.

The mechanism by which HIV-1 reverse transcriptase (HIV-RT) discriminates between the correct and incorrect nucleotide is not clearly understood. Chemically modified nucleotides containing 1-aminonaphthalene-5-sulfonate (ANS) attached to their {gamma}-phosphate were synthesized and used to probe nucleotide selection by this error prone polymerase. Primer extension reactions provide direct evidence that the polymerase is able to incorporate the gamma-modified nucleotides. Forward mutation assays reveal a 6-fold reduction in the mutational frequency with the modified nucleotides, and specific base substitutions are dramatically reduced or eliminated. Molecular modeling illustrates potential interactions between critical residues within the polymerase active site and the modified nucleotides. Our data demonstrate that the fidelity of reverse transcriptase is improved using modified nucleotides, and we suggest that specific modifications to the {gamma}-phosphate may be useful in designing new antiviral therapeutics or, more generally, as a tool for defining the structural role that the polymerase active site has on nucleotide selectivity.


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


This article has been cited by other articles:


Home page
 ScienceHome page
J. Eid, A. Fehr, J. Gray, K. Luong, J. Lyle, G. Otto, P. Peluso, D. Rank, P. Baybayan, B. Bettman, et al.
Real-Time DNA Sequencing from Single Polymerase Molecules
Science, January 2, 2009; 323(5910): 133 - 138.
[Abstract] [Full Text] [PDF]


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.