Overlapping signals for translational regulation and packaging of influenza A virus segment 2.

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 segment-specific packaging.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Wise HM,Barbezange C,Jagger BW,Dalton RM,Gog JR,Curran MD,Taubenberger JK,Anderson EC,Digard P

doi

10.1093/nar/gkr487

subject

Has Abstract

pub_date

2011-09-01 00:00:00

pages

7775-90

issue

17

eissn

0305-1048

issn

1362-4962

pii

gkr487

journal_volume

39

pub_type

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