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PMID: 2114979 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Eukaryotic coupled translation of tandem cistrons: identification of the influenza B virus BM2 polypeptide.

The EMBO journal ·Vol. 9 ·No. 8 ·1990-08-00 ·Pages 2639-47

Horvath CM, Williams MA, Lamb RA

Abstract

Previous nucleotide sequence analysis of RNA segment 7 of influenza B virus indicated that, in addition to the reading frame encoding the 248 amino acid M1 protein, there is a second overlapping reading frame (BM2ORF) of 585 nucleotides that has the coding capacity for 195 amino acids. To search for a polypeptide product derived from BM2ORF, a genetically engineered beta-galactosidase-BM2ORF fusion protein was expressed in Escherichia coli and a polyclonal rabbit antiserum was raised to the purified fusion protein. This antiserum was used to identify a polypeptide, designated BM2 protein (Mr approximately equal to 12,000), that is synthesized in influenza B virus-infected cells. To understand the mechanism by which the BM2 protein is generated from influenza B virus RNA segment 7, a mutational analysis of the cloned DNA was performed and the altered DNAs were expressed in eukaryotic cells. The expression patterns of the M1 and BM2 proteins from the altered DNAs indicate that the BM2 protein initiation codon overlaps with the termination codon of the M1 protein in an overlapping translational stop-start pentanucleotide, TAATG, and that the expression of the BM2 protein requires 5'-adjacent termination of M1 synthesis. Our data suggest that a termination-reinitiation scheme is used in translation of a bicistronic mRNA derived from influenza B virus RNA segment 7, and this strategy has some analogy to prokaryotic coupled stop-start translation of tandem cistrons.

MeSH Terms
Animals Base Sequence Cell Line Gene Expression Regulation, Viral Genes, Viral Influenza B virus/genetics Molecular Sequence Data Mutation Oligonucleotide Probes Plasmids Protein Biosynthesis RNA, Viral/genetics Recombinant Fusion Proteins/biosynthesis Restriction Mapping Transcription, Genetic Transfection Viral Proteins/genetics Viral Structural Proteins/genetics beta-Galactosidase/biosynthesis,genetics
Chemicals
BM2 polypeptide, influenza B virus Oligonucleotide Probes RNA, Viral Recombinant Fusion Proteins Viral Proteins Viral Structural Proteins beta-Galactosidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Horvath C M
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208-3500.
Williams M A
Lamb R A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-08-00
Pages
2639-47
Language
English
Region
England
NLM ID
8208664
PMCID
PMC552297
Subset
IM
Grants
NIAID NIH HHS · AI-20201 · United States
NIGMS NIH HHS · GM 08061 · United States
External Links
PubMed source PMC Full Text View journal details
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