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PMID: 2993644 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of cytomegalovirus gene expression: alpha and beta promoters are trans activated by viral functions in permissive human fibroblasts.

Journal of virology ·Vol. 56 ·No. 1 ·1985-10-00 ·Pages 135-43

Spaete RR, Mocarski ES

Abstract

We have fused immediate (alpha) and delayed (beta) early promoter-regulatory sequences taken from the cytomegalovirus (CMV) genome to Escherichia coli lacZ (beta-galactosidase) as an indicator gene to study regulated expression of these promoters. After transfection of human fibroblast cells with plasmid constructs carrying beta-galactosidase fusions, and subsequent infection with CMV, we have demonstrated that viral trans-acting functions up-regulate the expression of these genes in a temporally authentic manner. The alpha promoter is activated even when de novo protein synthesis is blocked and when UV-inactivated virus is used, suggesting that, as for herpes simplex virus type 1 (HSV-1), a virion structural protein is responsible for its up-regulation. We have found that HSV-1, as well as CMV, is capable of trans activating the CMV alpha promoter. The beta promoter is activated by CMV but is completely unresponsive to HSV-1 infection. The temporal synthesis of the alpha and beta promoters in the transient expression system conforms with their natural regulation during viral replication. The beta-galactosidase fusions we describe provide a most exquisitely sensitive indicator system for the study of cis- and trans-acting viral regulatory functions.

MeSH Terms
Cells, Cultured Cytomegalovirus/genetics DNA, Recombinant Gene Expression Regulation Genes, Viral Humans Promoter Regions, Genetic Protein Biosynthesis Simplexvirus/genetics Time Factors Virus Replication beta-Galactosidase/genetics
Chemicals
DNA, Recombinant beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Spaete R R
Mocarski E S
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53 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.56.1.135-143.1985
Published
1985-10-00
Pages
135-43
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC252498
Subset
IM
Grants
NIAID NIH HHS · 1 ROI AI20211 · United States
External Links
PubMed source DOI: 10.1128/JVI.56.1.135-143.1985 PMC Full Text View journal details
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