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PMID: 1651417 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Internal initiation of translation in retroviral vectors carrying picornavirus 5' nontranslated regions.

Journal of virology ·Vol. 65 ·No. 9 ·1991-09-00 ·Pages 4985-90

Adam MA, Ramesh N, Miller AD, Osborne WR

Abstract

Previous work has shown that picornavirus 5' nontranslated regions (NTRs) can initiate internal translation of downstream coding regions both in vitro and in transient in vivo assays. We have used 5' NTR sequences from encephalomyocarditis virus and poliovirus to construct retroviral vectors that are designed to express two proteins from a single mRNA. Inclusion of 5' NTR sequences did not adversely affect vector titer. Protein expression was studied with stable cell lines generated by vector infection of mouse NIH 3T3 cells and human and canine skin fibroblasts. Expression of a coding region in the downstream position was at levels from 25 to 100% of the same coding region in the upstream position. Expression of downstream coding regions in control vectors that did not contain the 5' NTR sequences was very low, in agreement with the predictions of the scanning model for eukaryotic translation. These experiments demonstrate coordinate expression of two coding regions from a single mRNA in stable cell lines and provide further support for the model of internal translation initiation by sequences in the 5' NTRs of picornaviruses.

Related Genes
ADA PNP
MeSH Terms
Adenosine Deaminase/genetics Animals Cells, Cultured Cloning, Molecular Dogs Encephalomyocarditis virus/genetics Gene Expression Genetic Vectors Humans Peptide Chain Initiation, Translational Poliovirus/genetics Polyribonucleotide Nucleotidyltransferase/genetics Regulatory Sequences, Nucleic Acid Retroviridae/genetics Virus Replication
Chemicals
Polyribonucleotide Nucleotidyltransferase Adenosine Deaminase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Adam M A
Program in Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Ramesh N
Miller A D
Osborne W R
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27 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.65.9.4985-4990.1991
Published
1991-09-00
Pages
4985-90
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC248961
Subset
IM
Grants
NIAID NIH HHS · AI19565 · United States
NHLBI NIH HHS · HL36444 · United States
NHLBI NIH HHS · HL41212 · United States
External Links
PubMed source DOI: 10.1128/JVI.65.9.4985-4990.1991 PMC Full Text View journal details
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