3'-end processing and kinetics of 5'-end joining during retroviral integration in vivo.

AUTOR(ES)

Roe, T

RESUMO

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.

Documentos Relacionados

• Alternative 3′-end processing of U5 snRNA by RNase III
• Transcription Termination and 3′-End Processing of the Spliced Leader RNA in Kinetoplastids
• Simian virus 40 minichromosomes as targets for retroviral integration in vivo.
• Nonhomologous End Joining during Restriction Enzyme-Mediated DNA Integration in Saccharomyces cerevisiae
• Independent 5' and 3'-end determination of multiple dihydrofolate reductase transcripts.