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Nucleic Acids Research Advance Access published online on July 18, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn474
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

The conserved N-terminal basic residues and zinc-finger motifs of HIV-1 nucleocapsid restrict the viral cDNA synthesis during virus formation and maturation

Ludovic Didierlaurent1,2,3, Laurent Houzet1,2,3, Zakia Morichaud1,2,3, Jean-Luc Darlix4 and Marylène Mougel1,2,3,*

1Université Montpellier 1 CPBS, 2CNRS, UMR 5236, CPBS, 4 Bd Henri IV, CS69033, 34965 Montpellier, 3Université Montpellier 2 and 4LaboRetro, Unité de virologie humaine INSERM U758, IFR128, ENS, Lyon, France

*To whom correspondence should be addressed. Tel: +33 4 67 60 02 32; Fax: +33 4 67 60 44 20; Email: mmougel{at}univ-montp1.fr

Received June 12, 2008. Revised July 5, 2008. Accepted July 7, 2008.

Reverse transcription of the genomic RNA by reverse transcriptase occurs soon after HIV-1 infection of target cells. The viral nucleocapsid (NC) protein chaperones this process via its nucleic acid annealing activities and its interactions with the reverse transcriptase enzyme. To function, NC needs its two conserved zinc fingers and flanking basic residues. We recently reported a new role for NC, whereby it negatively controls reverse transcription in the course of virus formation. Indeed, deleting its zinc fingers causes reverse transcription activation in virus producer cells. To investigate this new NC function, we used viruses with subtle mutations in the conserved zinc fingers and its flanking domains. We monitored by quantitative PCR the HIV-1 DNA content in producer cells and in produced virions. Results showed that the two intact zinc-finger structures are required for the temporal control of reverse transcription by NC throughout the virus replication cycle. The N-terminal basic residues also contributed to this new role of NC, while Pro-31 residue between the zinc fingers and Lys-59 in the C-terminal region did not. These findings further highlight the importance of NC as a major target for anti-HIV-1 drugs.

Present address: Laurent Houzet, LMM, NIAID, NIH, Bethesda, MD, USA


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