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Papers In Press, published online ahead of print November 30, 2001
J. Biol. Chem, 10.1074/jbc.M110830200
Submitted on November 12, 2001
Revised on November 30, 2001
Accepted on November 30, 2001

Ku represses the HIV-1 transcription: Identification of a putative Ku binding site homologous to the MMTV NRE1 sequence in the HIV-1 LTR

Laurence Jeanson and Jean-François Mouscadet

CNRS UMR 8532, Institut Gustave Roussy, Villejuif 94805

Corresponding Author: jfm{at}igr.fr

Ku has been implicated in nuclear processes including DNA break repair, transcription, V(D)J recombination and telomere maintenance. Its mode of action involves two distinct mechanisms; one in which a non specific binding occurs to DNA ends and a second that involves a specific binding to negative regulatory elements involved in transcription repression. Such elements were identified in MMTV and HTLV retroviruses. The purpose of this study was to investigate a role for Ku in the regulation of HIV-1 transcription. First, HIV-1 LTR activity was studied in CHO-K1 cells and in CH0-derived xrs-6 cells which are devoid of Ku80. LTR-driven expression of a reporter gene was significantly increased in xrs-6 cells. This enhancement was suppressed after re-expression of Ku80. Second, transcription of HIV-1 was followed in U1 human cells that were depleted in Ku by using a Ku80 antisense RNA. Ku depletion led to a increase of both HIV-1 mRNA synthesis and viral production compared to the parent cells. These results demonstrate that Ku acts as a transcriptional repressor of HIV-1 expression. Finally, a putative Ku specific binding site was identified within the negative regulatory region of the HIV-1 LTR which may account for this repression of transcription.


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