XL-MS of of influenza A NEP/NS2

ABSTRACT: The influenza nuclear export protein (NEP)/non-structural protein 2 (NS2) is a multifunctional protein, involved in viral ribonucleoprotein (vRNP) export from the nucleus, genome replication enhancement, and the adaption of avian influenza to mammals. Despite the growing attention on the importance of NEP in the influenza lifecycle and in interspecies transmission from avian hosts, the molecular details of how it is able to perform its various roles is still yet to be fully understood. In the interest of generating antiviral proteins and to aid in structural characterization, a panel of Rep proteins were raised against influenza virus A NEPH1N1. When complexed with RepE4 we were able to crystallize full-length NEPH1N1, revealing a folded and monomeric conformation. The N- and C-termini of NEPH1N1 pack together, with the middle linker region resembles a hinge, con-trasting a previous structure where NEP is dimeric and elongated. Together these structures demonstrate the plasticity of NEP, a trait which may potentially aid NEP in binding a diversity of cellular and viral partners. Using isothermal titration calorimetry (ITC) we measured a nanomolar interaction between RepE4 and NEP. Similarly, we found that RepE4 can also bind NEP from hu-man infecting avian H7N9 and bovine originating avian H5N1 influenza viruses. Owing to their high degree of conservation, RepE4 likely has the capacity to interact with NEP from numerous influenza A virus strains. Indeed, this, combined with the nanomolar affinity measured between NEP/RepE4 could be explored further as a broad-range therapeutic strategy and/or a tool in cellulo to understand NEP function at the molecular level.

INSTRUMENT(S):

ORGANISM(S): Influenza A Virus (a/philippines/tmc10-144/2010(h1n1))

SUBMITTER: Pascal Albanese  

LAB HEAD: Pascal Albanese

PROVIDER: PXD068009 | Pride | 2026-02-12

REPOSITORIES: Pride