Studies on the mechanism of replication of adenovirus DNA. II. The nature of single-stranded DNA in replicative intermediates.
Replicative intermediates of adenovirus type 5 DNA contain large stretches of single-stranded DNA. We have shown that this single-stranded DNA is mainly of parental origin, whereas all new DNA synthesized during one round of replication has a double-stranded structure. Hybridization experiments of the single-stranded DNA with isolated complementary strands of adenovirus type 5 DNA showed that this DNA hybridized only with the viral L-strand (the strand with the lower equilibrium density in alkaline CsCl) indicating that it represents the viral H-strand. This observation implies that replication always starts from one and the same molecular end. Electron microscopy of partially denatured Y-shaped intermediates confirmed this and showed that replication started from the molecular right end (the end richest in A-T base pairs). In conclusion, we have shown that replication of adenovirus type 5 DNA starts at the molecular right end, displacing the parental H-strand.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000256:Adenoviridae / Q000254:growth & development / Q000378:metabolism*; D002277:Carcinoma; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002845:Chromatography; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000096:biosynthesis*; D004279:DNA, Viral / Q000096:biosynthesis* / Q000302:isolation & purification; D006801:Humans; D008854:Microscopy, Electron; D009062:Mouth Neoplasms; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes; D013395:Sucrose; D014316:Tritium; D014779:Virus Replication
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D010761:Phosphorus Radioisotopes; D014316:Tritium; D002586:Cesium; D013395:Sucrose; D000255:Adenosine Triphosphate