Random-choice replication of extrachromosomal bovine papillomavirus (BPV) molecules in heterogeneous, clonally derived BPV-infected cell lines.
Using fluorescence in situ hybridization and Southern blot analysis, we show that three clonally derived cell lines transformed with bovine papillomavirus (BPV), including ID13, the cell line commonly employed for BPV replication studies, are heterogeneous populations having extensive cell-to-cell variation in both the distribution and amount of BPV DNA. Different subclones of ID13 were found to differ in the form and amount of BPV DNA they contain. Most subclones showed no detectable BPV sequences; some contained either extrachromosomal BPV molecules distributed throughout the nucleus or BPV sequences integrated at discrete chromosomal sites, while others contained both integrated and plasmid forms. The results of density gradient analysis of BPV DNA from individual homogeneous subclones showed replication of the extrachromosomal BPV plasmids in a random-choice mode. In all cell lines studied, the presence after one round of chromosomal DNA replication of unreplicated BPV DNA and of BPV DNA having two postreplicative strands was independent of the presence of high-BPV-copy-number ("jackpot") cells. Our results substantiate the earlier conclusion that extrachromosomal BPV molecules replicate randomly and not according to a once-per-cell-cycle mechanism.
Ravnan
J B
JB
Department of Genetics, Stanford University School of Medicine, California 94305-5120.
Gilbert
D M
DM
Ten Hagen
K G
KG
Cohen
S N
SN
D000818:Animals; D015139:Blotting, Southern; D052688:Bovine papillomavirus 1 / Q000502:physiology*; D002461:Cell Line, Transformed; D002499:Centrifugation, Density Gradient; D002999:Clone Cells; D004247:DNA / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D017403:In Situ Hybridization; D008938:Mitosis; D014779:Virus Replication
D004279:DNA, Viral; D004247:DNA