JBC Focus on PI3-Kinase with Echelon

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

A more recent version of this article appeared on August 8, 2003
This Article
Right arrow Full Text (Accepted Manuscript)
Right arrow All Versions of this Article:
278/32/29913    most recent
M211754200v1
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 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 Diamond, T. L.
Right arrow Articles by Kim, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Diamond, T. L.
Right arrow Articles by Kim, B.
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?

Papers In Press, published online ahead of print May 8, 2003
J. Biol. Chem, 10.1074/jbc.M211754200
Submitted on November 18, 2002
Revised on May 2, 2003
Accepted on May 8, 2003

Mechanistic understanding of an altered fidelity simian immunodeficiency virus reverse transcriptase mutation, V148I, identified in a pig-tailed macaque

Tracy L. Diamond, George Souroullas, Kellie K. Weiss, Kwi Y. Lee, Robert A. Bambara, Stephen Dewhurst, and Baek Kim

Microbiology and Immunology, University of Rochester, Rochester, NY 14642

Corresponding Author: baek_kim{at}urmc.rochester.edu

We have recently reported that the reverse transcriptase (RT) of SIVMNE 170 (170), which is a representative viral clone of the late symptomatic phase of infection with the parental strain, SIVMNE CL8 (CL8), has a largely increased fidelity, compared to the CL8 RT. In the present study, we analyzed the mechanistic alterations of the high fidelity 170 RT variant. Firstly, we found that among several 170 RT mutations, only one, V148I, is solely responsible for the fidelity increase over the CL8 RT. This V148I mutation lies near the Q151 residue that we recently found is important to the low fidelity of RT and the binding of incoming dNTPs. Therefore, secondly, we compared dNTP binding affinity (Kd) and catalysis (kpol) of the CL8 RT and the CL8-V148I RT using pre-steady state kinetic analysis. In this experiment, the high fidelity CL8-V148I RT has largely decreased binding to both correct and incorrect dNTP without altering kpol. The fidelity increase imparted by the V148I mutation is likely due to the major reduction seen in RT binding to dNTPs. This parallels our findings with the Q151N mutant. Thirdly, site-directed mutagenesis targeting amino acid residue 148 has revealed that a valine amino acid at this position is essential to RT infidelity. Based on these findings, we discuss possible structural impacts of residue 148 (and mutations at this site) on the interaction of RT with incoming dNTPs and infer how alterations in these properties may relate to viral replication and fitness.


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
V. K Jamburuthugoda, J. M. Santos-Velazquez, M. Skasko, D. J. Operario, V. Purohit, P. Chugh, E. A. Szymanski, J. E. Wedekind, R. A. Bambara, and B. Kim
Reduced dNTP Binding Affinity of 3TC-resistant M184I HIV-1 Reverse Transcriptase Variants Responsible for Viral Infection Failure in Macrophage
J. Biol. Chem., April 4, 2008; 283(14): 9206 - 9216.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. J. Operario, M. Balakrishnan, R. A. Bambara, and B. Kim
Reduced dNTP Interaction of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Promotes Strand Transfer
J. Biol. Chem., October 27, 2006; 281(43): 32113 - 32121.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
R. A. Smith, D. J. Anderson, and B. D. Preston
Hypersusceptibility to Substrate Analogs Conferred by Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase
J. Virol., July 15, 2006; 80(14): 7169 - 7178.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
V. K. Jamburuthugoda, P. Chugh, and B. Kim
Modification of Human Immunodeficiency Virus Type 1 Reverse Transcriptase to Target Cells with Elevated Cellular dNTP Concentrations
J. Biol. Chem., May 12, 2006; 281(19): 13388 - 13395.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Skasko, K. K. Weiss, H. M. Reynolds, V. Jamburuthugoda, K. Lee, and B. Kim
Mechanistic Differences in RNA-dependent DNA Polymerization and Fidelity between Murine Leukemia Virus and HIV-1 Reverse Transcriptases
J. Biol. Chem., April 1, 2005; 280(13): 12190 - 12200.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. L. Diamond, M. Roshal, V. K. Jamburuthugoda, H. M. Reynolds, A. R. Merriam, K. Y. Lee, M. Balakrishnan, R. A. Bambara, V. Planelles, S. Dewhurst, et al.
Macrophage Tropism of HIV-1 Depends on Efficient Cellular dNTP Utilization by Reverse Transcriptase
J. Biol. Chem., December 3, 2004; 279(49): 51545 - 51553.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.