Effect of Ku80 depletion on the preintegrative steps of HIV-1 replication in human cells.
To gain new insights regarding the role of Ku, the DNA-PK DNA-binding component, during lentiviral DNA integration, we have investigated the HIV-1 replication in Ku80-depleted human cells. CEM4fx cells underexpressing the Ku80 factor were selected after transduction by a retroviral vector expressing the Ku80 full-length antisense sequence. De novo infection experiment with NL4.3 HIV-1 strain led to the observation that the viral replication was delayed in the Ku80-depleted cells. Early events of the replicative cycle, including nuclear import of the viral DNA, were not affected. In contrast, the formation of the 2-LTR circles was impaired, thus demonstrating the implication of Ku in HIV-1 DNA circularization, for the first time in human cells. Furthermore, the detection of integrated proviruses by an Alu-LTR-nested PCR amplification method was affected in cells underexpressing Ku80. These results suggest that this factor may also be involved in the mechanisms leading to the stable establishment of HIV-1 provirus.
UMR8532 CNRS, Institut Gustave Roussy, PR2, 39 rue Camille Desmoulins, 94805, Villejuif, France.
Laurence
L
Jeanson
Frédéric
F
Subra
Sabine
S
Vaganay
Martial
M
Hervy
Elizabeth
E
Marangoni
Jean
J
Bourhis
Jean-François
JF
Mouscadet
D034961:Antigens, Nuclear; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004265:DNA Helicases; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D000072200:Ku Autoantigen; D009687:Nuclear Proteins / Q000172:deficiency / Q000378:metabolism*; D021381:Protein Transport; D014157:Transcription Factors / Q000172:deficiency / Q000378:metabolism; D016662:Virus Integration / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen