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PMID: 6319743 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

DNAs of two molecularly cloned endogenous ecotropic proviruses are poorly infectious in DNA transfection assays.

Journal of virology ·Vol. 49 ·No. 2 ·1984-02-00 ·Pages 437-44

Copeland NG, Bedigian HG, Thomas CY, Jenkins NA

Abstract

Endogenous ecotropic murine leukemia virus expression varies with inbred mouse strain and age. The mechanism(s) regulating virus expression is unknown, but expression is thought to be controlled at the transcriptional level by linkage to cis-acting cellular DNA sequences or DNA methylation or both. To begin to differentiate between these different control mechanisms, we molecularly cloned two endogenous ecotropic proviruses, Emv-3 and Emv-13, complete with flanking cellular DNA sequences. Both proviruses are poorly expressed in vivo and in vitro, although they appear to be structurally nondefective by restriction enzyme analysis. Cloned DNAs of both proviruses were poorly infectious in DNA transfection experiments, suggesting that methylation may not regulate the expression of these genes in vivo. Removal of their flanking cellular sequences did not increase their infectivity. However, these DNAs were highly infectious when mixed together, indicating that both proviruses carry mutations, that inhibit their expression and belong to different complementation groups. Marker rescue experiments suggested that Emv-3 is defective in the gag region and Emv-13 is defective in p15E-U3. The infectivity of Emv-3, but not of Emv-13, DNA was increased by the addition of AKR xenotropic murine leukemia virus DNA, consistent with known regions of homology between ecotropic and xenotropic proviruses. Recombination between defective endogenous viruses also appears to occur in vivo, suggesting that this may be a common mechanism controlling endogenous murine leukemia virus expression.

MeSH Terms
Animals Base Sequence Cells, Cultured Cloning, Molecular DNA Restriction Enzymes DNA, Viral/genetics Genes, Viral Leukemia Virus, Murine/genetics Mice Mutation Nucleic Acid Hybridization Transfection
Chemicals
DNA, Viral DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Copeland N G
Bedigian H G
Thomas C Y
Jenkins N A
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47 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.49.2.437-444.1984
Published
1984-02-00
Pages
437-44
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC255484
Subset
IM
Grants
NCI NIH HHS · CA-31102 · United States
NCI NIH HHS · CA-32324 · United States
NCI NIH HHS · CA-32995 · United States
External Links
PubMed source DOI: 10.1128/JVI.49.2.437-444.1984 PMC Full Text View journal details
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