Document Detail

Herpes simplex virus infection selectively stimulates accumulation of beta interferon reporter gene mRNA by a posttranscriptional mechanism.
MedLine Citation:
PMID:  1316484     Owner:  NLM     Status:  MEDLINE    
To study the mechanism of a novel herpes simplex virus (HSV) activity that stimulates expression of reporter genes containing beta interferon (IFN-beta)-coding sequences, we have established permanent DNA-transfected cell lines that each contain two distinct hybrid genes encoding mRNA species with different half-lives. These reporter genes comprised either the human IFN-beta- or bacterial chloramphenicol acetyltransferase (CAT)-coding and 3' untranslated regions placed under the transcriptional control of the powerful major immediate-early promoter-enhancer region (IE94) from simian cytomegalovirus. Most of the dual-transfected cell lines yielded significant levels of steady-state IE94-CAT mRNA and abundant constitutive synthesis of CAT enzyme activity, whereas no accumulation of IE94-IFN mRNA could be detected. However, infection with HSV type 1 resulted in a 300-fold increase in IE94-IFN-specific mRNA transcripts, compared with no more than 3- to 5-fold stimulation of IE94-CAT-specific mRNA. In contrast, cycloheximide treatment increased stable mRNA levels and transcription initiation rates from both the IE94-IFN and IE94-CAT hybrid genes. Run-on transcription assays in isolated nuclei suggested that induction of IE94-IFN gene expression by HSV type 1 occurred predominantly at the posttranscriptional level. Enhancement of the unstable IFN mRNA species after HSV infection was also observed in cell lines containing a simian virus 40 enhancer-driven IFN gene (SV2-IFN). Similarly, in transient-transfection assays, both SV2-IFN and IE94-IFN gave only low basal mRNA synthesis, but superinfection with HSV again led to high-level accumulation of IFN mRNA. Finally, substitution of the SV2-IFN gene 3' region with poly(A) and splicing signals from the SV2-CAT gene cassette led to stabilization of the IFN mRNA even in the absence of HSV. Therefore, we conclude that HSV infection leads to selective accumulation of IFN-beta mRNA by a posttranscriptional mechanism that is reporter gene specific and promoter independent.
J D Mosca; P M Pitha; G S Hayward
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of virology     Volume:  66     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  1992 Jun 
Date Detail:
Created Date:  1992-06-16     Completed Date:  1992-06-16     Revised Date:  2010-09-07    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  3811-22     Citation Subset:  IM    
Oncology Center, Johns Hopkins School of Medicine, Baltimore, Maryland 21205.
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MeSH Terms
Chloramphenicol O-Acetyltransferase / biosynthesis,  genetics
Cycloheximide / pharmacology
Cytomegalovirus / genetics
DNA, Recombinant / genetics
Herpes Simplex / metabolism*
Interferon-beta / biosynthesis,  genetics
Nucleic Acid Hybridization
RNA Probes
RNA, Messenger / metabolism*
Regulatory Sequences, Nucleic Acid / genetics*
Simplexvirus / genetics*
Transcription, Genetic* / drug effects
Transformation, Genetic
Grant Support
Reg. No./Substance:
0/DNA, Recombinant; 0/RNA Probes; 0/RNA, Messenger; 66-81-9/Cycloheximide; 77238-31-4/Interferon-beta; EC O-Acetyltransferase

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