A U5 repressor of reverse transcription is required for optimal HIV-1 infectivity and replication

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Title A U5 repressor of reverse transcription is required for optimal HIV-1 infectivity and replication
Author Meredith, Luke William; Ducloux, Céline; Isel, Catherine; Marquet, Roland; Harrich, David
Publication Title Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts
Year Published 2009
Place of publication United Kingdom
Publisher BioMed Central Ltd.
Abstract Here we provide strong evidence that a highly conserved stem loop structure in the U5 region of the HIV-1 RNA leader harbours a repressor of reverse transcription (RRT). We showed that two sequences in U5, at +143-145 and +151-153, are essential for RRT function. Mutation of either site strongly and unexpectedly increased endogenous reverse transcription, and cell infection assays showed that both mutations dramatically increased negative strand strong stop DNA synthesis. Early, late, 1-LTR and 2-LTR reverse transcription products were present proportionally, indicating that the downstream reverse transcription events were not affected. In vitro structural probing of the wild type and mutant RNA revealed an unexpected destabilization effect of the mutations on the whole U5 stem loop, which would explain the loss of regulation of reverse transcription. This functional effect was not observed in vitro, where, in the absence of viral proteins other than RT and cellular factors, all RNA performed similarly. These U5 mutations decreased virus replication in Jurkat and primary T-cells, which could be attributed to a marked defect in viral integration. Analysis of 1-LTR and 2-LTR circular DNA isolated from infected cells revealed that substantial deletions were present, indicating that the viral DNA was degraded by cellular nucleases. Together, our experiments suggest that regulated reverse transcription initiation is essential to allow synthesis of the viral DNA in a cellular environment that supports the assembly of a functional HIV-1 pre-integration complex, which also protects the proviral DNA from cellular degradation processes.
Peer Reviewed No
Published Yes
Publisher URI http://www.retrovirology.com/supplements/6/S2
Alternative URI http://dx.doi.org/10.1186/1742-4690-6-S2-O14
Copyright Statement Copyright 2009 Meredith et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ISBN 1742-4690
Conference name Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts
Location Montpellier, France
Date From 2009-09-21
Date To 2009-09-23
URI http://hdl.handle.net/10072/31543
Date Accessioned 2010-04-28
Language en_US
Faculty Griffith Health Faculty
Subject Microbiology
Publication Type Conference Publications (Extract Paper)
Publication Type Code e3

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