Deakin University
Browse

File(s) under permanent embargo

Expression and characterisation of the influenza A virus non-structural protein NS1 in yeast

Version 2 2024-06-03, 21:28
Version 1 2017-05-16, 15:53
journal contribution
posted on 2024-06-03, 21:28 authored by Alister WardAlister Ward, AA Azad, IG Macreadie
The influenza A virus non-structural protein NS1 was produced using a copper-inducible expression system in the yeast Saccharomyces cerevisiae. The protein produced had a molecular weight of 26 kDa by SDS-PAGE and was reactive with anti-NS1 antisera. The recombinant NS1 protein was targetted to the nucleolus/nuclear envelope fraction of the yeast cell nucleus, showing that its localisation signals remain functional in yeast. In addition, immune-electron microscopy detected cytoplasmic inclusions reminiscent of those seen in cells infected with some influenza strains. The NS1 protein was shown to be capable of in vivo self-interaction which probably forms the basis of its propensity to form inclusions. Expression of the protein was found to be toxic to yeast cells expressing it, supporting a role for the protein in the shutdown of influenza virus-infected cells. Deletion mapping of NS1 pointed to 2 regions of the molecule being important for this toxicity: a basic C-terminal stretch which has been shown to act as a nuclear localisation signal, and an N-terminal region implicated in RNA binding.

History

Journal

Archives of Virology

Volume

138

Pagination

299-314

Location

Wien, Austria

ISSN

0304-8608

Language

eng

Publication classification

C1.1 Refereed article in a scholarly journal

Copyright notice

1994, Springer-Verlag

Issue

3-4

Publisher

Springer Wien