Impact of the Ku complex on HIV-1 expression and latency.
Ku, a cellular complex required for human cell survival and involved in double strand break DNA repair and multiple other cellular processes, may modulate retroviral multiplication, although the precise mechanism through which it acts is still controversial. Recently, Ku was identified as a possible anti-human immunodeficiency virus type 1 (HIV-1) target in human cells, in two global approaches. Here we investigated the role of Ku on the HIV-1 replication cycle by analyzing the expression level of a panel of non-replicative lentiviral vectors expressing the green fluorescent protein in human colorectal carcinoma HCT 116 cells, stably or transiently depleted of Ku. We found that in this cellular model the depletion of Ku did not affect the efficiency of (pre-)integrative steps but decreased the early HIV-1 expression by acting at the transcriptional level. This negative effect was specific of the HIV-1 promoter, required the obligatory step of viral DNA integration and was reversed by transient depletion of p53. We also provided evidence on a direct binding of Ku to HIV-1 LTR in transduced cells. Ku not only promotes the early transcription from the HIV-1 promoter, but also limits the constitution of viral latency. Moreover, in the presence of a normal level of Ku, HIV-1 expression was gradually lost over time, likely due to the counter-selection of HIV-1-expressing cells. On the contrary, the reactivation of transgene expression from HIV-1 by means of trichostatin A- or tumor necrosis factor α-administration was enhanced under condition of Ku haplodepletion, suggesting a phenomenon of provirus latency. These observations plead in favor of the hypothesis that Ku has an impact on HIV-1 expression and latency at early- and mid-time after integration.
Laboratoire de Biologie et Pharmacologie Appliquée, Centre national de la recherche scientifique-UMR8113, Ecole Normale Supérieure de Cachan, Cachan, France.
Gwenola
G
Manic
Aurélie
A
Maurin-Marlin
Fanny
F
Laurent
Ilio
I
Vitale
Sylvain
S
Thierry
Olivier
O
Delelis
Philippe
P
Dessen
Michelle
M
Vincendeau
Christine
C
Leib-Mösch
Uriel
U
Hazan
Jean-François
JF
Mouscadet
Stéphanie
S
Bury-Moné
D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D045325:HCT116 Cells; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D000072200:Ku Autoantigen; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D016662:Virus Integration / Q000502:physiology; D017735:Virus Latency / Q000502:physiology
D016159:Tumor Suppressor Protein p53; D004265:DNA Helicases; C106893:XRCC5 protein, human; D000072200:Ku Autoantigen