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PMID: 2542958 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Initiation of simian virus 40 DNA replication in vitro: pulse-chase experiments identify the first labeled species as topologically unwound.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 11 ·1989-06-00 ·Pages 3944-8

Bullock PA, Seo YS, Hurwitz J

Abstract

A distinct unwound form of DNA containing the simian virus 40 (SV40) origin is produced in replication reactions carried out in mixtures containing crude fractions prepared from HeLa cells. This species, termed form UR, comigrates on chloroquine-containing agarose gels with the upper part of the previously described heterogeneous highly unwound circular DNA, form U. As with form U, formation of form UR is dependent upon the SV40 tumor (T) antigen. Pulse-chase experiments demonstrate that the first species to incorporate labeled deoxyribonucleotides comigrates with form UR. Restriction analyses of the products of the pulse-chase experiments show that initiation occurs at the SV40 origin and then proceeds outward in a bidirectional manner. These experiments establish form UR as the earliest detectable substrate for SV40 DNA replication and suggest that SV40 DNA replication initiates on an unwound species.

MeSH Terms
Animals Cell Line DNA Replication DNA, Viral/biosynthesis,genetics HeLa Cells/metabolism Humans Kinetics Nucleic Acid Conformation Restriction Mapping Simian virus 40/genetics
Chemicals
DNA, Viral
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bullock P A
Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Seo Y S
Hurwitz J
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36 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.11.3944
Published
1989-06-00
Pages
3944-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC287364
Subset
IM
Grants
NIGMS NIH HHS · GM 34559-06 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.11.3944 PMC Full Text View journal details
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