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PMID: 2153218 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Genetic analysis of the enhancer requirements for polyomavirus DNA replication in mice.

Journal of virology ·Vol. 64 ·No. 2 ·1990-02-00 ·Pages 476-85

Rochford R, Campbell BA, Villarreal LP

Abstract

In this report, we describe the first systematic analysis of the genetic requirements for polyomavirus (Py) enhancer-activated viral DNA replication during the acute phase of infection in mice. Four mutants were made which substituted XhoI sites for conserved enhancer consensus sequences (adenovirus type 5 E1A, c-fos, simian virus 40, and a glucocorticoidlike consensus sequence). Viral DNA replication in infected mouse organs was measured by DNA blot analysis. Only the loss of the glucocorticoidlike consensus sequence element significantly reduced Py DNA replication in the kidneys, the primary target organ for viral replication. The loss of the c-fos, adenovirus type 5 E1A, or simian virus 40 consensus sequences, however, expanded organ-specific viral DNA replication, relative to wild-type Py, by allowing high-level replication in the pancreas or heart or both. Analysis of Py variants selected for replication in undifferentiated embryonal carcinoma cell lines (PyF441, PyF111) showed that there was little change in levels of viral DNA replication in kidneys and other organs as compared with those in the wild-type virus. If the entire B enhancer is deleted, only low overall levels of viral replication are observed. Wild-type levels of replication in the kidneys can be reconstituted by addition of a single domain from within the A enhancer (nucleotides 5094 to 5132) to the B enhancer deletion virus, suggesting that a single domain from the A enhancer can functionally substitute for the entire B enhancer. This also indicates that the determinants for kidney-specific replication are not found in the B enhancer.

MeSH Terms
Animals Base Sequence Cell Line Chromosome Deletion Cloning, Molecular DNA Replication DNA, Viral/genetics,isolation & purification Enhancer Elements, Genetic Genes, Viral Genetic Variation Mice Mice, Inbred BALB C Molecular Sequence Data Oligonucleotide Probes Polyomavirus/genetics,pathogenicity Transfection Tumor Virus Infections/microbiology
Chemicals
DNA, Viral Oligonucleotide Probes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Rochford R
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Campbell B A
Villarreal L P
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.2.476-485.1990
Published
1990-02-00
Pages
476-85
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249134
Subset
IM
Grants
NIGMS NIH HHS · GM36605 · United States
External Links
PubMed source DOI: 10.1128/JVI.64.2.476-485.1990 PMC Full Text View journal details
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