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PMID: 2550797 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Origin auxiliary sequences can facilitate initiation of simian virus 40 DNA replication in vitro as they do in vivo.

Molecular and cellular biology ·Vol. 9 ·No. 9 ·1989-09-00 ·Pages 3593-602

Guo ZS, Gutierrez C, Heine U, Sogo JM, Depamphilis ML

Abstract

Initiation of simian virus 40 (SV40) DNA replication is facilitated by two auxiliary sequences that flank the minimally required origin (ori) core sequence. In monkey cells, the replication rate of each of the four ori configurations changed with time after transfection in a characteristic pattern. This pattern was reproduced in an extract from SV40-infected monkey cells by varying the ratio of DNA substrate to cell extract; DNA replication in vitro depended on ori auxiliary sequences to the same extent as they did in vivo. Facilitation by ori auxiliary sequences was lost at high ratios of DNA to cell extract, revealing that the activity of these sequences required either multiple initiation factors or a molar excess of one initiation factor bound to ori. This parameter, together with ionic strength and the method used to measure DNA replication, determined the level of facilitation by ori auxiliary sequences in vitro. The activity of ori auxiliary sequences was not diminished in vivo or in vitro by increasing amounts of large tumor antigen. Therefore, ori auxiliary sequences promoted initiation of replication at some step after tumor antigen binding to ori. Furthermore, although cellular factors could modulate the activity of ori auxiliary sequences in vitro, these factors did not appear to involve nucleosome assembly because no correlation was observed between the number of nucleosomes assembled per DNA molecule and facilitation by ori auxiliary sequences. These results demonstrate that SV40 ori auxiliary sequences can function in vitro as they do in vivo and begin to elucidate their role in initiating DNA replication.

MeSH Terms
Animals Antigens, Viral, Tumor Base Sequence Cell Line, Transformed Chromosome Mapping DNA Replication DNA, Viral/genetics Genes, Viral Humans Plasmids Simian virus 40/genetics,immunology,physiology Virus Replication
Chemicals
Antigens, Viral, Tumor DNA, Viral
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Guo Z S
Department of Cell and Developmental Biology, Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
Gutierrez C
Heine U
Sogo J M
Depamphilis M L
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.9.3593-3602.1989
Published
1989-09-00
Pages
3593-602
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362419
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.9.9.3593-3602.1989 PMC Full Text View journal details
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