Nucleic Acids Research Advance Access originally published online on November 7, 2006
Nucleic Acids Research 2006 34(21):6215-6224; doi:10.1093/nar/gkl843
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Nucleic Acids Research, 2006, Vol. 34, No. 21 6215-6224
© 2006 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 |
Chromosomal integration of LTR-flanked DNA in yeast expressing HIV-1 integrase: down regulation by RAD51
1 UMR 5097-CNRS, Bordeaux France 2 Université Victor Segalen Bordeaux 2, Bordeaux France 3 IFR 66 ‘Pathologies Infectieuses et Cancers’, Bordeaux France 4 146 rue Léo Saignat, 33076 Bordeaux cedex France 5 Deptartment of Molecular Biophysics and Biochemistry, Yale University School of Medicine 333 Cedar Street, SHM C130, New Haven, CT 06520, USA
*To whom correspondence should be addressed. Tel: +33 5 57 57 1740; Fax: +33 5 57 57 1766; Email: vincent.parissi{at}reger.u-bordeaurd.fr
Received June 7, 2006. Revised October 3, 2006. Accepted October 9, 2006.
HIV-1 integrase (IN) is the key enzyme catalyzing the proviral DNA integration step. Although the enzyme catalyzes the integration step accurately in vitro, whether IN is sufficient for in vivo integration and how it interacts with the cellular machinery remains unclear. We set up a yeast cellular integration system where integrase was expressed as the sole HIV-1 protein and targeted the chromosomes. In this simple eukaryotic model, integrase is necessary and sufficient for the insertion of a DNA containing viral LTRs into the genome, thereby allowing the study of the isolated integration step independently of other viral mechanisms. Furthermore, the yeast system was used to identify cellular mechanisms involved in the integration step and allowed us to show the role of homologous recombination systems. We demonstrated physical interactions between HIV-1 IN and RAD51 protein and showed that HIV-1 integrase activity could be inhibited both in the cell and in vitro by RAD51 protein. Our data allowed the identification of RAD51 as a novel in vitro IN cofactor able to down regulate the activity of this retroviral enzyme, thereby acting as a potential cellular restriction factor to HIV infection.