Replication process of the parvovirus H-1. II. Isolation and characterization of H-1 replicative form DNA.
The Parvovirus H-1 replicates autonomously in hamster embryo cells. A DNA synthetic event, called HA-DNA synthesis, upon which subsequent viral RNA and viral hemagglutinin synthesis is dependent, is initiated in late S phase of the infected cell (18). It was postulated that HA-DNA represents parental viral replicative form DNA (RF DNA). This study describes the isolation and characterization of H-1 RF DNA as part of the continuing study of the mechanisms and control of DNA replication in the eukaryotic cell. The H-1 RF DNA is a linear duplex molecule containing the viral strand and its complement. The complementary strands of the RF DNA have been separated by equilibrium density gradient centrifugation. The RF DNA has a buoyant density of 1.705 in neutral CsCl and an estimated guanine plus cytosine (GC) content of 45.9%. It has a sedimentation coefficient of 17S. The calculated molecular weight of 3.7 x 10(6) is twice that of the single-stranded virion DNA. H-1 virions contain DNA that is homogeneous and free of complementary strands.
D000818:Animals; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D006224:Cricetinae; D003596:Cytosine / Q000032:analysis; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D006147:Guanine / Q000032:analysis; D006882:Hydroxyapatites; D008722:Methods; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010321:Parvoviridae / Q000032:analysis / Q000254:growth & development* / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D014779:Virus Replication
D002250:Carbon Radioisotopes; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D006882:Hydroxyapatites; D014316:Tritium; D006147:Guanine; D003596:Cytosine; D013936:Thymidine