Identification of putative interactions between swine and human influenza A virus nucleoprotein and human host proteins

Abstract

BackgroundInfluenza A viruses (IAVs) are important pathogens that affect the health of humans and many additional animal species. IAVs are enveloped, negative single-stranded RNA viruses whose genome encodes at least ten proteins. The IAV nucleoprotein (NP) is a structural protein that associates with the viral RNA and is essential for virus replication. Understanding how IAVs interact with host proteins is essential for elucidating all of the required processes for viral replication, restrictions in species host range, and potential targets for antiviral therapies.MethodsIn this study, the NP from a swine IAV was cloned into a yeast two-hybrid “bait” vector for expression of a yeast Gal4 binding domain (BD)-NP fusion protein. This “bait” was used to screen a Y2H human HeLa cell “prey” library which consisted of human proteins fused to the Gal4 protein’s activation domain (AD). The interaction of “bait” and “prey” proteins resulted in activation of reporter genes.ResultsSeventeen positive bait-prey interactions were isolated in yeast. All of the “prey” isolated also interact in yeast with a NP “bait” cloned from a human IAV strain. Isolation and sequence analysis of the cDNAs encoding the human prey proteins revealed ten different human proteins. These host proteins are involved in various host cell processes and structures, including purine biosynthesis (PAICS), metabolism (ACOT13), proteasome (PA28B), DNA-binding (MSANTD3), cytoskeleton (CKAP5), potassium channel formation (KCTD9), zinc transporter function (SLC30A9), Na+/K+ ATPase function (ATP1B1), and RNA splicing (TRA2B).ConclusionsTen human proteins were identified as interacting with IAV NP in a Y2H screen. Some of these human proteins were reported in previous screens aimed at elucidating host proteins relevant to specific viral life cycle processes such as replication. This study extends previous findings by suggesting a mechanism by which these host proteins associate with the IAV, i.e., physical interaction with NP. Furthermore, this study revealed novel host protein-NP interactions in yeast.

Influenza A Virus-induced expression of ISG20 inhibits viral replication by interacting with nucleoprotein

“…NP is a multi-functional protein of IAV, and several host factors were identified to interact with NP, and the interaction between NP and host factors exhibited its diverse role in viral life cycle [37][38][39][40][41][42]. The human anti-apoptosis protein clusterin (CLU) is targeted by NP to induced apoptosis [41].…”

Prediction of non-canonical routes for SARS-CoV-2 infection in human placenta cells

“…We found that PAICS transcript, which is also translated into a protein predicted as interacting with the Nsp3 of SARS-CoV, decreased its levels with placental development and growth. PAICS is an enzyme of de novo purine biosynthesis pathway, which was previously predicted as having a putative interaction with the human influenza A virus (IAV) nucleoprotein and was upregulated during IAV infection (34). The putative interaction of Nsp3 with PAICS is relevant.…”

Structural Insights into KCTD Protein Assembly and Cullin3 Recognition

“…This results in upregulated CD69 expression, cytotoxicity, interferon-secretion, and downregulation of the NKG2A receptor [27][28][29]. A recent study has also shown KCTD9 may interact with influenza A proteins [30]. The natural biological role of KCTD9 in the absence of viral infection, and the function of KCTD9/Cul3 binding in viral liver injury, remains unknown.…”