Abstract
Influenza A viruses (IAV) are enveloped viruses carrying a single-stranded negative-sense RNA genome. Detection of host proteins having a relationship with IAV and revealing of the role of these proteins in the viral replication are of great importance in keeping IAV infections under control. Consequently, the importance of human DDX56, which is determined to be associated with a viral NS1 with a yeast two-hybrid assay, was investigated for IAV replication. The viral replication in knocked down cells for the DDX56 gene was evaluated. The NS1 was co-precipitated with the DDX56 protein in lysates of cells transiently expressing DDX56 and NS1 or infected with the viruses, showing that NS1 and DDX56 interact in mammalian cells. Viral NS1 showed a tendency to co-localize with DDX56 in the cells, transiently expressing both of these proteins, which supports the IP and two-hybrid assays results. The data obtained with in silico predictions supported the in vitro protein interaction results. The viral replication was significantly reduced in the DDX56-knockdown cells comparing with that in the control cells. In conclusion, human DDX56 protein interacts with the IAV NS1 protein in both yeast and mammalian cells and has a positive regulatory effect on IAV replication. However, the mechanism of DDX56 on IAV replication requires further elucidation.
Highlights
Influenza A viruses are enveloped viruses with a singlestranded RNA genome having a negative-polarity
The cDNAs were belong to genes coding DEAD (Asp-Glu-Ala-Asp) box helicase 56 (DDX56), neuroguidin, EIF4E binding protein (NGDN), proteosome subunit beta type 4 (PSMB4), ribosomal protein L29 (RPL29), and small nuclear ribonucleoprotein D1 polypeptide (SNRPD1) (Figures S1-S5)
Influenza A virus RNA dependent RNA polymerase (RdRP) removes the 5′ cap structure of pre-mRNAs synthesized by RNA pol II and uses it in the transcription of viral mRNAs (Dias et al, 2009)
Summary
Influenza A viruses are enveloped viruses with a singlestranded RNA genome having a negative-polarity. The NS2 protein, which is found in the lowest amount in the virion, has a role in the transport of viral ribonucleoprotein particles (RNPs) with M1 protein and several cellular proteins (Senbas Akyazi et al, 2020). The influenza A virus NS1 protein has an average of 26 kDa molecular weight and consists of 230 or 237 amino acid residues depending on the virus type (Lin et al, 2007). This protein has the functional domains for both protein-protein and protein-RNA interactions
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