Activation of PKR by Bunyamwera virus is independent of the viral interferon antagonist NSs.
Double-stranded RNA (dsRNA) is a by-product of viral RNA polymerase activity, and its recognition is one mechanism by which the innate immune system is activated. Cellular responses to dsRNA include induction of alpha/beta interferon (IFN) synthesis and activation of the enzyme PKR, which exerts its antiviral effect by phosphorylating the eukaryotic initiation factor eIF-2 alpha, thereby inhibiting translation. We have recently identified the nonstructural protein NSs of Bunyamwera virus (BUNV), the prototype of the family Bunyaviridae, as a virulence factor that blocks the induction of IFN by dsRNA. Here, we investigated the potential of NSs to inhibit PKR. We show that wild-type (wt) BUNV that expresses NSs triggered PKR-dependent phosphorylation of eIF-2 alpha to levels similar to those of a recombinant virus that does not express NSs (BUNdelNSs virus). Furthermore, the sensitivity of viruses in cell culture to IFN was independent of PKR and was not determined by NSs. PKR knockout mice, however, succumbed to infection approximately 1 day earlier than wt mice or mice deficient in expression of RNase L, another dsRNA-activated antiviral enzyme. Our data indicate that (i) bunyaviruses activate PKR, but are only marginally sensitive to its antiviral effect, and (ii) NSs is different from other IFN antagonists, since it inhibits dsRNA-dependent IFN induction but has no effect on the dsRNA-activated PKR and RNase L systems.
Streitenfeld
Hein
H
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, Germany.
Boyd
Amanda
A
Fazakerley
John K
JK
Bridgen
Anne
A
Elliott
Richard M
RM
Weber
Friedemann
F
D000818:Animals; D002042:Bunyamwera virus / Q000472:pathogenicity* / Q000502:physiology; D002044:Bunyaviridae Infections / Q000401:mortality / Q000821:virology; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004268:DNA-Binding Proteins / Q000378:metabolism; D004789:Enzyme Activation; D007372:Interferons / Q000037:antagonists & inhibitors*; D051379:Mice; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014774:Virulence
D004268:DNA-Binding Proteins; C103652:Elf2 protein, mouse; D014157:Transcription Factors; D017361:Viral Nonstructural Proteins; D007372:Interferons; D017346:Protein Serine-Threonine Kinases