Simian virus 40 minichromosomes as targets for retroviral integration in vivo.
We present a method for studying multiple retroviral integration events into a small DNA target in vivo. Episomal simian virus 40 (SV40) genomes established by infection of CV-1 cells served as integration targets during subsequent infection with murine leukemia virus (MLV). Using a PCR-based assay for the abundance and distribution of integration events, nonrandom integration of MLV DNA into SV40 DNA is detectable as early as 4 hr and reaches a maximum level by 8 hr after MLV infection. The level of integration but not the distribution of integration sites is sensitive to the stage in the SV40 life cycle at which MLV infection is performed. Using a temperature-sensitive tumor (T) antigen mutant SV40 strain, we observed that active replication of the target DNA is not required for efficient integration in vivo. The distribution of integration sites in vivo is closely approximately by in vitro reactions with isolated SV40 minichromosomes as integration targets. However, the degree of bias between the most and least favored sites is greater in vivo than in vitro.
Pryciak
P M
PM
Department of Biochemistry, University of California, San Francisco 94143-0502.
Müller
H P
HP
Varmus
H E
HE
D000818:Animals; D002460:Cell Line; D016679:Genome, Viral; D000882:Haplorhini; D007700:Kinetics; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013539:Simian virus 40 / Q000235:genetics* / Q000502:physiology; D013997:Time Factors; D014771:Virion / Q000235:genetics / Q000502:physiology; D016662:Virus Integration