3'-end processing and kinetics of 5'-end joining during retroviral integration in vivo.
Retroviral replication depends on integration of viral DNA into a host cell chromosome. Integration proceeds in three steps: 3'-end processing, the endonucleolytic removal of the two terminal nucleotides from each 3' end of the viral DNA; strand transfer, the joining of the 3' ends of viral DNA to host DNA; and 5'-end joining (or gap repair), the joining of the 5' ends of viral DNA to host DNA. The 5'-end joining step has never been investigated, either for retroviral integration or for any other transposition process. We have developed an assay for 5'-end joining in vivo and have examined the kinetics of 5'-end joining for Moloney murine leukemia virus (MLV). The interval between 3'-end and 5'-end joining is estimated to be less than 1 h. This assay will be a useful tool for examining whether viral or host components mediate 5'-end joining. MLV integrates its DNA only after its host cell has completed mitosis. We show that the extent of 3'-end processing is the same in unsynchronized and aphidicolin-arrested cells. 3'-end processing therefore does not depend on mitosis.
Roe
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Department of Biochemistry, Stanford University Medical Center, California 94305, USA.
Chow
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Brown
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D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D004279:DNA, Viral / Q000235:genetics*; D009052:Leukemia Virus, Murine / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D016662:Virus Integration / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral