Skip Navigation

This Article
Right arrow Print PDF (1770K)
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 Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (29)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by DeStefano, J. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by DeStefano, J. J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1995, Vol. 23, No. 19 3901-3908
© 1995

ENZYMOLOGY

The orientation of binding of human immunodeficiency virus reverse transcriptase on nucleic acid hybrids

Jeffrey J. DeStefano

Department of Microbiology Building 231, University of Maryland, College Park, MD 20742, USA

Received June 27, 1995. Revised August 22, 1995. Accepted August 22, 1995.

The binding of HIV reverse transcriptase (RT) to heteroduplexes was examined using a substrate consisting of a 42 nt chimeric nucleic acid composed (5'->3') of 23 nt of RNA and 19 of DNA. This chimera was hybridized to an internal region of a relatively long complementary DNA or RNA. When the chimera was bound to DNA and conditions limiting cleavage to a single binding event between the enzyme and sub strata were employed initial RNase H-directed cleavages occurred 19-21 nt from the chimera 5'-terminus. A 42 nt strand Identical in sequence to the chimera and composed of only RNA was cleaved at the same locations. Reducing the length of the DNA portion of the chlmera from 19 to 7 nt did not after the cleavage positions, suggesting that cleavage was not coordi nated by the DNA 3'-terminus. Under the same conditions cleavage was not detected when the chimera was bound to RNA. In contrast, addition of dNTPs to the DNA 3'-terminus of the chimera occurred only when the chimera was bound to RNA. The resuits support preferable binding of AT to RNA-DNA versus DNA-DNA hybrid regions and a model in which the orientation of binding to heteroduplexes is 5'->3' (relative to the RNA strand), poiymerase to ANase H active site, wfth sites associated with the DNA and RNA strand respectively.


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
 J. Biol. Chem.Home page
S. J. Schultz, M. Zhang, and J. J. Champoux
Sequence, Distance, and Accessibility Are Determinants of 5'-End-directed Cleavages by Retroviral RNases H
J. Biol. Chem., January 27, 2006; 281(4): 1943 - 1955.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. J. DeStefano and J. V. Cristofaro
Selection of primer-template sequences that bind human immunodeficiency virus reverse transcriptase with high affinity
Nucleic Acids Res., January 5, 2006; 34(1): 130 - 139.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
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]

Home page
 J. Virol.Home page
K. Post, J. Guo, K. J. Howard, M. D. Powell, J. T. Miller, A. Hizi, S. F. J. Le Grice, and J. G. Levin
Human Immunodeficiency Virus Type 2 Reverse Transcriptase Activity in Model Systems That Mimic Steps in Reverse Transcription
J. Virol., July 1, 2003; 77(13): 7623 - 7634.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
S. J. Schultz, M. Zhang, and J. J. Champoux
Specific Cleavages by RNase H Facilitate Initiation of Plus-Strand RNA Synthesis by Moloney Murine Leukemia Virus
J. Virol., May 1, 2003; 77(9): 5275 - 5285.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Raja and J. J. DeStefano
Interaction of HIV Reverse Transcriptase with Structures Mimicking Recombination Intermediates
J. Biol. Chem., March 14, 2003; 278(12): 10102 - 10111.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
C. S. Snyder and M. J. Roth
Comparison of Second-Strand Transfer Requirements and RNase H Cleavages Catalyzed by Human Immunodeficiency Virus Type 1 Reverse Transcriptase (RT) and E478Q RT
J. Virol., October 15, 2000; 74(20): 9668 - 9679.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
C. D. Kelleher and J. J. Champoux
RNA Degradation and Primer Selection by Moloney Murine Leukemia Virus Reverse Transcriptase Contribute to the Accuracy of Plus Strand Initiation
J. Biol. Chem., April 21, 2000; 275(17): 13061 - 13070.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. J. Schultz, M. Zhang, C. D. Kelleher, and J. J. Champoux
Polypurine Tract Primer Generation and Utilization by Moloney Murine Leukemia Virus Reverse Transcriptase
J. Biol. Chem., December 3, 1999; 274(49): 34547 - 34555.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. D. Powell, W. A. Beard, K. Bebenek, K. J. Howard, S. F. J. Le Grice, T. A. Darden, T. A. Kunkel, S. H. Wilson, and J. G. Levin
Residues in the alpha H and alpha I Helices of the HIV-1 Reverse Transcriptase Thumb Subdomain Required for the Specificity of RNase H-catalyzed Removal of the Polypurine Tract Primer
J. Biol. Chem., July 9, 1999; 274(28): 19885 - 19893.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Gotte, G. Maier, A. M. Onori, L. Cellai, M. A. Wainberg, and H. Heumann
Temporal Coordination between Initiation of HIV (+)-Strand DNA Synthesis and Primer Removal
J. Biol. Chem., April 16, 1999; 274(16): 11159 - 11169.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Palaniappan, J. K. Kim, M. Wisniewski, P. J. Fay, and R. A. Bambara
Control of Initiation of Viral Plus Strand DNA Synthesis by HIV Reverse Transcriptase
J. Biol. Chem., February 13, 1998; 273(7): 3808 - 3816.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. D. Powell, M. Ghosh, P. S. Jacques, K. J. Howard, S. F. J. Le Grice, and J. G. Levin
Alanine-scanning Mutations in the "Primer Grip" of p66 HIV-1 Reverse Transcriptase Result in Selective Loss of RNA Priming Activity
J. Biol. Chem., May 16, 1997; 272(20): 13262 - 13269.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Palaniappan, M. Wisniewski, P. S. Jacques, S. F.J. Le Grice, P. J. Fay, and R. A. Bambara
Mutations within the Primer Grip Region of HIV-1 Reverse Transcriptase Result in Loss of RNase H Function
J. Biol. Chem., April 25, 1997; 272(17): 11157 - 11164.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. J. Schultz, M. Zhang, C. D. Kelleher, and J. J. Champoux
Analysis of Plus-strand Primer Selection, Removal, and Reutilization by Retroviral Reverse Transcriptases
J. Biol. Chem., October 6, 2000; 275(41): 32299 - 32309.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Wisniewski, M. Balakrishnan, C. Palaniappan, P. J. Fay, and R. A. Bambara
The Sequential Mechanism of HIV Reverse Transcriptase RNase H
J. Biol. Chem., November 22, 2000; 275(48): 37664 - 37671.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. J. DeStefano, J. V. Cristofaro, S. Derebail, W. P. Bohlayer, and M. J. Fitzgerald-Heath
Physical Mapping of HIV Reverse Transcriptase to the 5' End of RNA Primers
J. Biol. Chem., August 24, 2001; 276(35): 32515 - 32521.
[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.