Isolation of the viral DNA replication complex from adeno-associated virus type 1-infected cells.
The replication complex active in adeno-associated virus type 1 (AAV-1) DNA synthesis in vitro was solubilized, with a nonionic detergent, from the nuclei of human embryonic kidney cells coinfected with AAV-1 and an early temperative-sensitive mutant (ts125) of human adenovirus type 5 at the nonpermissive temperature (40.5 degrees C). The complex sedimented with a mean size of 23S and contained parental AAV-1 DNA. Most of the DNA synthesized with the AAV-1 DNA replication complex in vitro was AAV-1 DNA, as revealed by DNA-DNA hybridization and sedimentation in a neutral sucrose gradient. However, it sedimented in an alkaline sucrose gradient as molecules smaller than AAV-1 DNA (14.4S). The AAV-1 DNA replication complex was not formed in cells infected with AAV-1 alone.
D000256:Adenoviridae; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004261:DNA Replication; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010321:Parvoviridae; D012526:Satellite Viruses; D014779:Virus Replication
D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase