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PMID: 1851878 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Herpes simplex virus origin-binding protein (UL9) loops and distorts the viral replication origin.

Journal of virology ·Vol. 65 ·No. 6 ·1991-06-00 ·Pages 3284-92

Koff A, Schwedes JF, Tegtmeyer P

Abstract

To investigate the role of the herpes simplex virus origin-binding protein (UL9) in the initiation of DNA replication, we have examined the effect of UL9 binding on the structure of the viral origin of replication. UL9 loops and alters the DNA helix of the origin regardless of the phasing of the binding sites. DNase I and micrococcal nuclease footprinting show that UL9 binds two sites in the origin and loops the AT-rich DNA between them independent of the topology of the DNA. KMnO4 and dimethyl sulfate footprinting further show that UL9 alters the DNA helix in the AT region. In contrast to the looping reaction, however, helical distortion requires the free energy of supercoiled DNA. UL9 also loops and distorts the origin DNA of a replication-defective mutant with a 6-bp insertion in the AT region. Because the helical distortion of this mutant DNA is different from that of functional origins, we conclude that an imperfect tertiary structure of the mutant DNA may contribute to its loss of replication function.

MeSH Terms
Animals Base Composition Base Sequence Binding Sites DNA Replication DNA, Superhelical/chemistry DNA, Viral/chemistry DNA-Binding Proteins/chemistry Deoxyribonuclease I Models, Molecular Molecular Sequence Data Moths/microbiology Mutation Nucleic Acid Conformation Protein Conformation Simplexvirus/growth & development Thermodynamics Viral Proteins/chemistry Virus Replication
Chemicals
DNA, Superhelical DNA, Viral DNA-Binding Proteins Viral Proteins origin-binding proteins, viral Deoxyribonuclease I
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Koff A
Department of Microbiology, State University of New York, Stony Brook 11794-8621.
Schwedes J F
Tegtmeyer P
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.65.6.3284-3292.1991
Published
1991-06-00
Pages
3284-92
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240986
Subset
IM
Grants
NCI NIH HHS · CA-09176 · United States
NCI NIH HHS · CA-18808 · United States
NCI NIH HHS · CA-28146 · United States
External Links
PubMed source DOI: 10.1128/JVI.65.6.3284-3292.1991 PMC Full Text View journal details
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