Identification of novel host cell binding partners of Oas1b, the protein conferring resistance to flavivirus-induced disease in mice

J Virol. 2012 Aug;86(15):7953-63. doi: 10.1128/JVI.00333-12. Epub 2012 May 23.

Abstract

Oas1b was previously identified as the product of the Flv(r) allele that confers flavivirus-specific resistance to virus-induced disease in mice by an uncharacterized, RNase L-independent mechanism. To gain insights about the mechanism by which Oas1b specifically reduces the efficiency of flavivirus replication, cellular protein interaction partners were identified and their involvement in the Oas1b-mediated flavivirus resistance mechanism was analyzed. Initial difficulties in getting the two-hybrid assay to work with full-length Oas1b led to the discovery that this Oas protein uniquely has a C-terminal transmembrane domain that targets it to the endoplasmic reticulum (ER). Two peptides matching to oxysterol binding protein-related protein 1L (ORP1L) and ATP binding cassette protein 3, subfamily F (ABCF3), were identified as Oas1b interaction partners in yeast two-hybrid assays, and both in vitro-transcribed/translated peptides and full-length proteins in mammalian cell lysates coimmunoprecipitated with Oas1b. Knockdown of a partner involved in Oas1b-mediated antiflavivirus activity would be expected to increase flavivirus replication but not that of other types of viruses. However, RNA interference (RNAi) knockdown of ORP1L decreased the replication of the flavivirus West Nile virus (WNV) as well as that of other types of RNA viruses. This virus-nonspecific effect may be due to the recently reported dysregulation of late endosome movement by ORP1L knockdown. Knockdown of ABCF3 protein levels increased the replication of WNV but not that of other types of RNA viruses, and this effect on WNV replication was observed only in Oas1b-expressing cells. The results suggest that Oas1b is part of a complex located in the ER and that ABCF3 is a component of the Flv(r)-mediated resistance mechanism.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2',5'-Oligoadenylate Synthetase / genetics
  • 2',5'-Oligoadenylate Synthetase / immunology
  • 2',5'-Oligoadenylate Synthetase / metabolism*
  • Animals
  • Cell Line
  • Cricetinae
  • Disease Resistance / physiology*
  • Endoplasmic Reticulum / genetics
  • Endoplasmic Reticulum / metabolism
  • Endoplasmic Reticulum / physiology
  • Endoplasmic Reticulum / virology
  • Flavivirus / physiology*
  • Flavivirus Infections / genetics
  • Flavivirus Infections / immunology
  • Flavivirus Infections / metabolism*
  • Gene Knockdown Techniques
  • Mice
  • Two-Hybrid System Techniques
  • Virus Replication / physiology*

Substances

  • Oas1b protein, mouse
  • 2',5'-Oligoadenylate Synthetase