Abstract
RNA secondary structures play a key role in splicing, gene expression, microRNA biogenesis, RNA editing, and other biological processes. The importance of RNA structures has been demonstrated in the life cycle of RNA-containing viruses, including the influenza virus. At least two regions of conserved secondary structure in NS segment (+) RNA are predicted to vary among influenza virus strains with respect to thermodynamic stability; both fall in the NS1 open reading frame. The NS1 protein is involved in multiple virus-host interaction processes, and its main function is to inhibit the cellular immune response to viral infection. Using a reverse genetics approach, four influenza virus strains were constructed featuring mutations that have different effects on RNA secondary structure. Growth curve experiments and ELISA data show that, at least in the first viral replication cycle, mutations G123A and A132G affecting RNA structure in the (82–148) NS RNA region influence NS1 protein expression.
Highlights
The influenza A virus (IAV) poses a serious threat to human health
IAV belongs to the Ortomyxoviridae family, and its genome consists of eight segments of negative sense RNA which encode more than 17 proteins [1]
RNA secondary structures at NS nucleotide regions 82– 148 and 497–564 have been previously predicted for a large number of influenza A viruses [12]
Summary
The influenza A virus (IAV) poses a serious threat to human health. Despite significant progress in surveillance and control measures, including the development of antiviral drugs, vaccines and diagnostics, IAV continues to evolve and cause epidemics. Other researchers have shown that mutant influenza A viruses with altered RNA structure in the second region display changes in NS mRNA splicing and viral replication in cell culture [16]. We analyzed the roles in NS1 protein expression and viral reproduction in vitro of mutations that influence conserved RNA secondary structures located in two NS regions.
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