Interferon antagonist NSs of La Crosse virus triggers a DNA damage response-like degradation of transcribing RNA polymerase II.
La Crosse encephalitis virus (LACV) is a mosquito-borne member of the negative-strand RNA virus family Bunyaviridae. We have previously shown that the virulence factor NSs of LACV is an efficient inhibitor of the antiviral type I interferon system. A recombinant virus unable to express NSs (rLACVdelNSs) strongly induced interferon transcription, whereas the corresponding wt virus (rLACV) suppressed it. Here, we show that interferon induction by rLACVdelNSs mainly occurs through the signaling pathway leading from the pattern recognition receptor RIG-I to the transcription factor IRF-3. NSs expressed by rLACV, however, acts downstream of IRF-3 by specifically blocking RNA polymerase II-dependent transcription. Further investigations revealed that NSs induces proteasomal degradation of the mammalian RNA polymerase II subunit RPB1. NSs thereby selectively targets RPB1 molecules of elongating RNA polymerase II complexes, the so-called IIo form. This phenotype has similarities to the cellular DNA damage response, and NSs was indeed found to transactivate the DNA damage response gene pak6. Moreover, NSs expressed by rLACV boosted serine 139 phosphorylation of histone H2A.X, one of the earliest cellular reactions to damaged DNA. However, other DNA damage response markers such as up-regulation and serine 15 phosphorylation of p53 or serine 1524 phosphorylation of BRCA1 were not triggered by LACV infection. Collectively, our data indicate that the strong suppression of interferon induction by LACV NSs is based on a shutdown of RNA polymerase II transcription and that NSs achieves this by exploiting parts of the cellular DNA damage response pathway to degrade IIo-borne RPB1 subunits.
Department of Virology, Institute for Medical Microbiology and Hygiene, University of Freiburg, D-79008 Freiburg, Germany.
Paul
P
Verbruggen
Marius
M
Ruf
Gjon
G
Blakqori
Anna K
AK
Överby
Martin
M
Heidemann
Dirk
D
Eick
Friedemann
F
Weber
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004249:DNA Damage; D004795:Enzyme Stability; D006801:Humans; D007372:Interferons / Q000037:antagonists & inhibitors; D018062:La Crosse virus / Q000472:pathogenicity*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000502:physiology*
D017361:Viral Nonstructural Proteins; D007372:Interferons; D012319:RNA Polymerase II