Title : Molecular characterization of nucleoporins–influenza A virus interactions: Potential host-directed therapeutic target
Abstract:
Influenza A virus (IAV), a negative-sense single-stranded RNA virus, primarily relies on the host nucleocytoplasmic transport machinery for trafficking of viral ribonucleoprotein (vRNP) complexes. The nuclear pore complex (NPC), composed of nucleoporins (NUPs), serves as a crucial interface exploited by viruses for molecular exchange between the nucleus and cytoplasm. In the present study, we investigated interactions between human nucleoporins and influenza A virus, integrating in-silico and experimental approaches. We have performed molecular docking to predict binding affinities at the protein–protein interface, and molecular dynamics simulations were carried out to evaluate the conformation and structural stability of the protein complexes under physiological conditions. Computational analyses revealed significant interactions between nucleoporins, namely NUP54 and NUP62, with the IAV vRNP protein throughout the 100 ns simulation period. Our results indicated that the NUPs interact with the influenza virus nucleoprotein via diverse mechanisms, with the NUP54-NP complex exhibiting dynamic rearrangements, whereas NUP62-NP remains stable. Subsequently, in vitro studies were performed to validate the functional relevance of these interactions. Increased NUP54 expression was observed with viral NP accumulation at the early stages of infection, suggesting an active role for NUP54 during viral replication. Notably, siRNA-mediated silencing of NUP shows approximately 60-70% reduction in viral infection, indicating that NUP54 contributes substantially to vRNP trafficking. This study provides novel insights into the role of NPC-associated proteins during influenza A virus replication. The identification of conserved host factors involved in virus infection highlights their potential for antiviral therapeutics to overcome rapid antigenic drift and antiviral resistance. Our findings provide a framework for functional characterization of host-virus mechanisms that may contribute to the development of effective host-directed vaccine strategies.
