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J. Biol. Chem., Vol. 277, Issue 7, 4918-4924, February 15, 2002

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Ku Represses the HIV-1 Transcription
IDENTIFICATION OF A PUTATIVE Ku BINDING SITE HOMOLOGOUS TO THE MOUSE MAMMARY TUMOR VIRUS NRE1 SEQUENCE IN THE HIV-1 LONG TERMINAL REPEAT^*

Laurence Jeanson and Jean-François Mouscadet

From the CNRS UMR8532, Institut Gustave-Roussy, PR2, 39 rue Camille Desmoulins, 94805 Villejuif, France

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 nonspecific 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 mouse mammary tumor virus and human T cell leukemia virus retroviruses. The purpose of this study was to investigate a role for Ku in the regulation of human immunodeficiency virus (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 with 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 long terminal repeat, which may account for this repression of transcription.

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