The Replication Pattern of Adenovirus DNA in vivo Reproduced in vitro

• Published in: European journal of biochemistry Vol. 104; no. 2; pp. 587 - 594
• Main Authors: CHEN, Shih‐min, ZUBAY, Geoffrey, GINSBERG, Harold S.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.03.1980
• Subjects: Adenoviruses, Human - metabolism; Carcinoma; Cell Line; Cell-Free System; DNA Replication; DNA, Viral - biosynthesis; Humans; Kinetics; Mouth Neoplasms; Nucleic Acid Hybridization; Virus Replication
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1980.tb04462.x

Temperature‐sensitive mutants of adenovirus type 5 (H5ts125 and H5ts149), which are conditionally inhibited in the initiation of viral DNA synthesis, have been exploited to investigate the possibility of the initiation of replication in a cell‐free system. Nuclei were isolated from human KB cells which had been infected with wild‐type or mutant adenovirus. More than 90 % of the DNA synthesis taking place in such nuclei was virus‐specific and the pattern of drug inhibition suggested that the synthesis required DNA polymerase γ. Nuclei prepared from cells infected with the H5ts125 temperature‐sensitive mutant which have been shifted from 33 °C to 39.5 °C showed a pattern of synthesis in vitro which began at both ends of the viral genome and gradually spread through the rest of the molecule.