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PMID: 2168979 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Herpes simplex virus DNA synthesis at a preformed replication fork in vitro.

Journal of virology ·Vol. 64 ·No. 10 ·1990-10-00 ·Pages 4957-67

Rabkin SD, Hanlon B

Abstract

Proteins from herpes simplex virus (HSV)-infected cells were used to reconstitute DNA synthesis in vitro on a preformed replication fork. The preformed replication fork consisted of a nicked, double-stranded, circular DNA molecule with a 5' single-strand tail that was noncomplementary to the template. The products of DNA synthesis on this substrate were rolling-circle molecules, as demonstrated by electron microscopy and alkaline agarose gel electrophoresis. The tails contained double-stranded regions, indicating that both leading- and lagging-strand DNA syntheses occurred. Rolling-circle DNA replication was dependent upon HSV DNA polymerase and ATP and was stimulated by a crude fraction containing ICP8 (HSV DNA-binding protein). Similar protein fractions from mock-infected cells were unable to support rolling-circle DNA replication. This in vitro DNA replication system should prove useful in the identification and characterization of the enzymatic activities required at the HSV replication fork.

MeSH Terms
Acyclovir/analogs & derivatives,pharmacology Animals Antiviral Agents/pharmacology Cell Nucleus/metabolism Cell Transformation, Viral DNA Replication/drug effects DNA, Viral/biosynthesis,isolation & purification,ultrastructure DNA-Directed DNA Polymerase/isolation & purification,metabolism Kinetics Microscopy, Electron Oligonucleotide Probes Simplexvirus/drug effects,enzymology,genetics Vero Cells
Chemicals
Antiviral Agents DNA, Viral Oligonucleotide Probes acyclovir triphosphate DNA-Directed DNA Polymerase Acyclovir
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rabkin S D
Program in Molecular Biology, Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
Hanlon B
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.10.4957-4967.1990
Published
1990-10-00
Pages
4957-67
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC247987
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.64.10.4957-4967.1990 PMC Full Text View journal details
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