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

Curt M. Horvath*, Mark A. Williams, Robert A. Lamb

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

111 Scopus citations

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 Ml 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 β-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 ≈ 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 Ml and BM2 proteins from the altered DNAs indicate that the BM2 protein initiation codon overlaps with the termination codon of the Ml 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.

Original languageEnglish (US)
Pages (from-to)2639-2647
Number of pages9
JournalEMBO Journal
Volume9
Issue number8
StatePublished - 1990

Keywords

  • BM2 protein
  • Overlapping reading frames
  • Translation stop - Start

ASJC Scopus subject areas

  • General Immunology and Microbiology
  • General Biochemistry, Genetics and Molecular Biology
  • Molecular Biology
  • General Neuroscience

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