CRK adaptor protein expression is required for efficient replication of avian influenza A viruses and controls JNK-mediated apoptotic responses

Cell Microbiol. 2010 Jun;12(6):831-43. doi: 10.1111/j.1462-5822.2010.01436.x. Epub 2010 Jan 29.

Abstract

The non-structural protein 1 (A/NS1) of influenza A viruses (IAV) harbours several src-homology domain (SH) binding motifs that are required for interaction with cellular proteins. The SH3 binding motif at aa212-217 [PPLPPK] of A/NS1 was shown to be essential for binding to the cellular adaptor proteins CRK and CRKL. Both regulate diverse cellular effector pathways, including activation of the MAP-kinase JNK that in turn mediates antiviral responses to IAV infection. By studying functional consequences of A/NS1-CRK interaction we show here that A/NS1 binding to CRK contributes to suppression of the antiviral-acting JNK-ATF2 pathway. However, only IAV that encode an A/NS1-protein harbouring the CRK/CRKL SH3 binding motif PPLPPK were attenuated upon downregulation of CRKI/II and CRKL, but not of CRKII alone. The PPLPPK site-harbouring candidate strains could be discriminated from other strains by a pronounced viral activation of the JNK-ATF2 signalling module that was even further boosted upon knock-down of CRKI/II. Interestingly, this enhanced JNK activation did not alter type-I IFN-expression, but rather resulted in increased levels of virus-induced cell death. Our results imply that binding capacity of A/NS1 to CRK/CRKL has evolved in virus strains that over-induce the antiviral acting JNK-ATF2 signalling module and helps to suppress the detrimental apoptosis promoting action of this pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 2
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Apoptosis*
  • Cell Line
  • Chick Embryo
  • Dogs
  • Humans
  • Influenza A virus / growth & development*
  • JNK Mitogen-Activated Protein Kinases / metabolism*
  • Nuclear Proteins / metabolism
  • Protein Binding
  • Proto-Oncogene Proteins c-crk / metabolism*
  • Viral Nonstructural Proteins / metabolism*

Substances

  • ATF2 protein, human
  • Activating Transcription Factor 2
  • Adaptor Proteins, Signal Transducing
  • CRK protein, human
  • CRKL protein
  • INS1 protein, influenza virus
  • Nuclear Proteins
  • Proto-Oncogene Proteins c-crk
  • Viral Nonstructural Proteins
  • JNK Mitogen-Activated Protein Kinases