Wise, Helen M., Barbezange, Cyril, Jagger, Brett W., Dalton, Rosa Maria, Gog, Julia R., Curran, Martin D., Taubenberger, Jeffery, Anderson, Emma C. and Digard, Paul. (2011) Overlapping signals for translational regulation and packaging of influenza A virus segment 2. Nucleic Acids Research, Vol.39 (No.17). pp. 7775-7790. ISSN 1362-4962

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Abstract

Influenza A virus segment 2 mRNA expresses three polypeptides: PB1, PB1-F2 and PB1-N40, from AUGs 1, 4 and 5 respectively. Two short open reading frames (sORFs) initiated by AUGs 2 and 3 are also present. To understand translational regulation in this system, we systematically mutated AUGs 1–4 and monitored polypeptide synthesis from plasmids and recombinant viruses. This identified sORF2 as a key regulatory element with opposing effects on PB1-F2 and PB1-N40 expression. We propose a model in which AUGs 1–4 are accessed by leaky ribosomal scanning, with sORF2 repressing synthesis of downstream PB1-F2. However, sORF2 also up-regulates PB1-N40 expression, most likely by a reinitiation mechanism that permits skipping of AUG4. Surprisingly, we also found that in contrast to plasmid-driven expression, viruses with improved AUG1 initiation contexts produced less PB1 in infected cells and replicated poorly, producing virions with elevated particle:PFU ratios. Analysis of the genome content of virus particles showed reduced packaging of the mutant segment 2 vRNAs. Overall, we conclude that segment 2 mRNA translation is regulated by a combination of leaky ribosomal scanning and reinitiation, and that the sequences surrounding the PB1 AUG codon are multifunctional, containing overlapping signals for translation initiation and for segmentspecific packaging.

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Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology > QR355 Virology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH):	Influenza A virus -- Genetics, Genetic translation
Journal or Publication Title:	Nucleic Acids Research
Publisher:	Oxford University Press
ISSN:	1362-4962
Date:	21 June 2011
Volume:	Vol.39
Number:	No.17
Page Range:	pp. 7775-7790
Identification Number:	10.1371/journal.pone.0023630
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Medical Research Council (Great Britain) (MRC), National Institutes of Health (U.S.) (NIH), Oxford/Cambridge Research Scholars Program, National Institute of Allergy and Infectious Diseases (U.S.) (NIAID)
Grant number:	G0700815 (MRC), G0701220 (MRC)
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URI:	http://wrap.warwick.ac.uk/id/eprint/38328

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