| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print June 11, 2003
Biochemistry and Biophysics, University of Rochester, Rochester, NY 14642
Corresponding Author: robert_bambara{at}urmc.rochester.edu
Template switching during reverse transcription promotes recombination in retroviruses. Efficient switches have been measured in vitro on hairpin-containing RNA templates, by a two-step mechanism. Pausing of the reverse transcriptase (RT) at the hairpin base allowed enhanced cleavage of the initial donor RNA template, exposing regions of the cDNA and allowing the acceptor to base pair with the cDNA. This defines the first or docking step. The primer continued synthesis on the donor, transferring or locking in a second step. Here we determine the enzyme dependant factors that influence template switching by comparing the RTs from HIV-1 and EIAV. HIV-1 RT promoted transfers with higher efficiency than EIAV RT. We found that both RTs paused strongly at the base of the hairpin. While stalled, HIV-1 RT made closely spaced cuts, while EIAV RT made only a single cut. Docking occurred efficiently at the multiply but not at the singly cut site. HIV-1 nucleocapsid protein (NC) stimulated strand transfers. It improved RNase H activity of both RTs. It allowed the EIAV RT to make a distribution of cuts, greatly stimulating docking at the base of the hairpin. Most likely, it also promoted strand exchange, allowing transfers to be initiated from sites throughout the hairpin. Minor pause sites beyond the base of the hairpin correlated with the locking sites. The strand exchange properties of NC likely promote this step. We present a model that explains the roles of RNase H specificity, template structure and properties of NC in the two-step transfer reaction.
J. Biol. Chem, 10.1074/jbc.M304608200
Submitted on May 2, 2003
Revised on June 4, 2003
Accepted on June 10, 2003
Role of the reverse transcriptase, nucleocapsid protein and template structure in the two step transfer mechanism in retroviral recombination
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Song, V. P. Basu, M. N. Hanson, B. P. Roques, and R. A. Bambara Proximity and Branch Migration Mechanisms in HIV-1 Minus Strand Strong Stop DNA Transfer J. Biol. Chem., February 8, 2008; 283(6): 3141 - 3150. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-Y. Li, F. Guo, L. Zhang, L. Kleiman, and S. Cen APOBEC3G Inhibits DNA Strand Transfer during HIV-1 Reverse Transcription J. Biol. Chem., November 2, 2007; 282(44): 32065 - 32074. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. D. Pratico and S. K. Silverman Ty1 reverse transcriptase does not read through the proposed 2',5'-branched retrotransposition intermediate in vitro RNA, September 1, 2007; 13(9): 1528 - 1536. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Wu, S. L. Heilman-Miller, and J. G. Levin Effects of nucleic acid local structure and magnesium ions on minus-strand transfer mediated by the nucleic acid chaperone activity of HIV-1 nucleocapsid protein Nucleic Acids Res., June 9, 2007; 35(12): 3974 - 3987. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Purohit, B. P. Roques, B. Kim, and R. A. Bambara Mechanisms That Prevent Template Inactivation by HIV-1 Reverse Transcriptase RNase H Cleavages J. Biol. Chem., April 27, 2007; 282(17): 12598 - 12609. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gao, M. Balakrishnan, B. P. Roques, and R. A. Bambara Insights into the Multiple Roles of Pausing in HIV-1 Reverse Transcriptase-promoted Strand Transfers J. Biol. Chem., March 2, 2007; 282(9): 6222 - 6231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
M. Song, M. Balakrishnan, Y. Chen, B. P. Roques, and R. A. Bambara Stimulation of HIV-1 Minus Strand Strong Stop DNA Transfer by Genomic Sequences 3' of the Primer Binding Site J. Biol. Chem., August 25, 2006; 281(34): 24227 - 24235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Lanciault and J. J. Champoux Pausing during Reverse Transcription Increases the Rate of Retroviral Recombination J. Virol., March 1, 2006; 80(5): 2483 - 2494. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Purohit, M. Balakrishnan, B. Kim, and R. A. Bambara Evidence That HIV-1 Reverse Transcriptase Employs the DNA 3' End-directed Primary/Secondary RNase H Cleavage Mechanism during Synthesis and Strand Transfer J. Biol. Chem., December 9, 2005; 280(49): 40534 - 40543. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Heras, M. C. Lopez, J. L. Garcia-Perez, S. L. Martin, and M. C. Thomas The L1Tc C-Terminal Domain from Trypanosoma cruzi Non-Long Terminal Repeat Retrotransposon Codes for a Protein That Bears Two C2H2 Zinc Finger Motifs and Is Endowed with Nucleic Acid Chaperone Activity Mol. Cell. Biol., November 1, 2005; 25(21): 9209 - 9220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Chen, M. Balakrishnan, B. P. Roques, and R. A. Bambara Acceptor RNA Cleavage Profile Supports an Invasion Mechanism for HIV-1 Minus Strand Transfer J. Biol. Chem., April 15, 2005; 280(15): 14443 - 14452. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. D. Rhodes, O. Nikolaitchik, J. Chen, D. Powell, and W.-S. Hu Genetic Recombination of Human Immunodeficiency Virus Type 1 in One Round of Viral Replication: Effects of Genetic Distance, Target Cells, Accessory Genes, and Lack of High Negative Interference in Crossover Events J. Virol., February 1, 2005; 79(3): 1666 - 1677. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Lanciault and J. J. Champoux Effects of Unpaired Nucleotides within HIV-1 Genomic Secondary Structures on Pausing and Strand Transfer J. Biol. Chem., January 28, 2005; 280(4): 2413 - 2423. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. S. Derebail and J. J. DeStefano Mechanistic Analysis of Pause Site-dependent and -independent Recombinogenic Strand Transfer from Structurally Diverse Regions of the HIV Genome J. Biol. Chem., November 12, 2004; 279(46): 47446 - 47454. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Heilman-Miller, T. Wu, and J. G. Levin Alteration of Nucleic Acid Structure and Stability Modulates the Efficiency of Minus-Strand Transfer Mediated by the HIV-1 Nucleocapsid Protein J. Biol. Chem., October 15, 2004; 279(42): 44154 - 44165. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
