Interaction of influenza A virus NS2/NEP protein with the amino-terminal part of Nup214.

Burçak Şenbaş Akyaz,Kadir Turan,Atsushi Kawaguchi,Ayşegül Pi̇ri̇nçal,Kyosuke Nagata

doi:10.3906/biy-1909-49

Burçak Şenbaş Akyaz, Kadir Turan + Show 3 more

Open Access

https://doi.org/10.3906/biy-1909-49

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Abstract

Influenza A viruses have a single-stranded RNA genome consisting of 8 segments. Each RNA segment associates with the nucleoprotein (NP) and viral RNA polymerase to and from a viral ribonucleoprotein (vRNP) particle. The viral mRNA synthesis is dependent on a capped primer derived from nascent host RNA transcripts. For these processes to take place, vRNPs must pass through the cell nuclear pore complex (NPC) to the nucleus. The influenza A virus NS2 protein, also called the nuclear export protein (NES), has an important role in the nucleocytoplasmic transport of vRNPs. This protein interacts with the host cellular nucleoporins during the nuclear export of vRNPs. In this study, the human nucleoporin 214 (Nup214) was identified as an NS2-binding protein by using a yeast two-hybrid assay. The interaction between NS2 and human Nup214 was confirmed in both yeast and mammalian cells. It has been shown that the NS2 protein interacts with the amino terminal FG domain of the Nup214 protein. The influenza viral replication was suppressed in knockdown cells for the Nup214 protein. It was concluded that the FG domains of nucleoporins have an important role in the interaction of the influenza NS2 protein with host NPC for vRNA export.

Highlights

Influenza A viruses are members of the family Orthomyxoviridae, which includes enveloped and segmented negative-sense single-stranded RNA viruses
Influenza A virus NS2 protein interacts with human nucleoporin 214 (Nup214) in yeast cells In order to identify the host proteins that interact with the influenza A virus NS2 protein, yeast two-hybrid screen experiments were performed by using a plasmid-coding NS2 bait and a human cDNA library cloned into pACT2
We screened the potential cellular proteins encoded from human cDNAs for interaction with the influenza virus NS2 protein by using a yeast twohybrid assay, and found an interaction between the NS2 and the Nup214 proteins

Summary

Introduction

Influenza A viruses are members of the family Orthomyxoviridae, which includes enveloped and segmented negative-sense single-stranded RNA (ssRNA–) viruses. The primary dependence of influenza virus transcription differentiates these viruses from most other RNA viruses which replicate in the cytoplasm. Upon infection of the host cell, influenza vRNPs transport into the nucleus, where both transcription and replication of each vRNA is carried out (Kemler et al, 1994). The viral RNA-dependent RNA polymerase (RdRp) enzyme uses the capped primer derived from host mRNA to initiate transcription (Lukarska et al, 2017; Plotch et al, 1981). In order for these processes to take place, vRNPs must pass

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