Formation of oligomeric free BK virus DNA is required for tandem integration of viral genomes into cellular DNA.
Mouse kidney cells have been transformed by linear BK virus (BKV) DNA with cohesive ends. These BKV genomes can circularize and subsequently replicate or physically join end to end yielding oligomeric viral DNA molecules. Blot hybridization analysis of transformed cells showed the presence of BKV DNA both in free circular forms and integrated into cellular DNA in a tandem head-to-tail array of full-length viral genomes. Formation of oligomers either by replication via previous circularization or by end to end joining is hindered after elimination of cohesive termini by digestion with the single-strand-specific nuclease S1. After treatment of linear BKV DNA with nuclease S1 only integrations of single viral genomes were observed in transformed cells; tandem insertions and free viral DNA were absent. These results support the hypothesis that formation of tandems of free viral DNA is a necessary prerequisite for tandem integration of viral genomes.
D000818:Animals; D001739:BK Virus / Q000235:genetics*; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D005814:Genes, Viral; D051379:Mice; D009693:Nucleic Acid Hybridization; D011120:Polyomavirus / Q000235:genetics*
D004279:DNA, Viral; D004247:DNA