HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fuentes, G. M. Articles by Bambara, R. A. Search for Related Content PubMed PubMed Citation Articles by Fuentes, G. M. Articles by Bambara, R. A. Social Bookmarking                      What's this?

Volume 271, Number 47, Issue of November 22, 1996 pp. 29605-29611
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Strand Displacement Synthesis in the Central Polypurine Tract Region of HIV-1 Promotes DNA to DNA Strand Transfer Recombination

(Received for publication, April 25, 1996)

Gloria M. Fuentes , Chockalingam Palaniappan ^§ , Philip J. Fay ^§¶ and Robert A. Bambara ^§

From the Departments of  Microbiology & Immunology, ^§ Biochemistry, and ^¶ Medicine, and the  Cancer Center, University of Rochester, School of Medicine and Dentistry, Rochester, New York 14642

Two distinct plus strand initiation sites have been identified in human immunodeficiency virus (HIV), the central polypurine tract (cPPT) and the polypurine tract located just upstream of the U3 region (U3-PPT). When synthesis from the U3-PPT reaches the cPPT, the elongating primer causes limited strand displacement of the product created from the cPPT. We examined whether reverse transcriptase (RT) catalyzed strand transfer recombination is promoted by this process. Using a substrate having the viral sequence of the displaced region, we measured transfer of an elongating DNA primer from a donor DNA to an acceptor DNA. Strand transfer synthesis was only efficient when RT was performing strand displacement synthesis. Transfer efficiency was directly related to acceptor concentration but independent of the reaction time. Transfer could occur to acceptors containing 80, 40, or 20 nucleotides of homology with the template DNA. Using different acceptors, we found that DNA to DNA transfer occurred at positions throughout the donor template, except near the 5 end. This shows that a number of the sequences downstream of the cPPT region can promote transfer, but once synthesis has progressed to the point where the downstream segment is completely displaced transfer is not allowed. When the DNA to DNA transfer reactions were performed using a template containing nonviral sequences, the transfer efficiency dropped significantly. This indicates that transfer efficiency is determined by the sequences of the templates used. HIV-RT RNase H-dependent strand transfer between RNA templates is well documented. We propose a quite different mechanism for DNA to DNA transfer, consistent with the ability of RNase H minus RT to perform this reaction. If these DNA to DNA transfer events occur in vivo, they will result in plus strand recombination.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				D. J. Garfinkel, K. M. Stefanisko, K. M. Nyswaner, S. P. Moore, J. Oh, and S. H. Hughes Retrotransposon Suicide: Formation of Ty1 Circles and Autointegration via a Central DNA Flap J. Virol., December 15, 2006; 80(24): 11920 - 11934. [Abstract] [Full Text] [PDF]

				C. Lanciault and J. J. Champoux Single Unpaired Nucleotides Facilitate HIV-1 Reverse Transcriptase Displacement Synthesis through Duplex RNA J. Biol. Chem., July 30, 2004; 279(31): 32252 - 32261. [Abstract] [Full Text] [PDF]

				S. Lyonnais, C. Hounsou, M.-P. Teulade-Fichou, J. Jeusset, E. L. Cam, and G. Mirambeau G-quartets assembly within a G-rich DNA flap. A possible event at the center of the HIV-1 genome Nucleic Acids Res., December 1, 2002; 30(23): 5276 - 5283. [Abstract] [Full Text] [PDF]

				L. Hameau, J. Jeusset, S. Lafosse, D. Coulaud, E. Delain, T. Unge, T. Restle, E. Le Cam, and G. Mirambeau Human Immunodeficiency Virus Type 1 Central DNA Flap: Dynamic Terminal Product of Plus-Strand Displacement DNA Synthesis Catalyzed by Reverse Transcriptase Assisted by Nucleocapsid Protein J. Virol., April 1, 2001; 75(7): 3301 - 3313. [Abstract] [Full Text]

				G. Dornadula, S. Yang, R. J. Pomerantz, and H. Zhang Partial Rescue of the Vif-Negative Phenotype of Mutant Human Immunodeficiency Virus Type 1 Strains from Nonpermissive Cells by Intravirion Reverse Transcription J. Virol., March 15, 2000; 74(6): 2594 - 2602. [Abstract] [Full Text]

				C. M. Smith, J. S. Smith, and M. J. Roth RNase H Requirements for the Second Strand Transfer Reaction of Human Immunodeficiency Virus Type 1 Reverse Transcription J. Virol., August 1, 1999; 73(8): 6573 - 6581. [Abstract] [Full Text]

				T. Wu, J. Guo, J. Bess, L. E. Henderson, and J. G. Levin Molecular Requirements for Human Immunodeficiency Virus Type 1 Plus-Strand Transfer: Analysis in Reconstituted and Endogenous Reverse Transcription Systems J. Virol., June 1, 1999; 73(6): 4794 - 4805. [Abstract] [Full Text]

				J. A. Rumbaugh, G. M. Fuentes, and R. A. Bambara Processing of an HIV Replication Intermediate by the Human DNA Replication Enzyme FEN1 J. Biol. Chem., October 30, 1998; 273(44): 28740 - 28745. [Abstract] [Full Text] [PDF]