The pathogenic NY-1 hantavirus G1 cytoplasmic tail inhibits RIG-I- and TBK-1-directed interferon responses.
Hantaviruses cause two diseases with prominent vascular permeability defects, hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome. All hantaviruses infect human endothelial cells, although it is unclear what differentiates pathogenic from nonpathogenic hantaviruses. We observed dramatic differences in interferon-specific transcriptional responses between pathogenic and nonpathogenic hantaviruses at 1 day postinfection, suggesting that hantavirus pathogenesis may in part be determined by viral regulation of cellular interferon responses. In contrast to pathogenic NY-1 virus (NY-1V) and Hantaan virus (HTNV), nonpathogenic Prospect Hill virus (PHV) elicits early interferon responses following infection of human endothelial cells. We determined that PHV replication is blocked in human endothelial cells and that RNA and protein synthesis by PHV, but not NY-1V or HTNV, is inhibited at 2 to 4 days postinfection. The addition of antibodies to beta interferon (IFN-beta) blocked interferon-directed MxA induction by >90% and demonstrated that hantavirus infection induces the secretion of IFN-beta from endothelial cells. Coinfecting endothelial cells with NY-1V and PHV resulted in a 60% decrease in the induction of interferon-responsive MxA transcripts by PHV and further suggested the potential for NY-1V to regulate early IFN responses. Expression of the NY-1V G1 cytoplasmic tail inhibited by >90% RIG-I- and downstream TBK-1-directed transcription from interferon-stimulated response elements or beta-interferon promoters in a dose-dependent manner. In contrast, expression of the NY-1V nucleocapsid or PHV G1 tail had no effect on RIG-I- or TBK-1-directed transcriptional responses. Further, neither the NY-1V nor PHV G1 tails inhibited transcriptional responses directed by a constitutively active form of interferon regulatory factor 3 (IRF-3 5D), and IRF-3 is a direct target of TBK-1 phosphorylation. These findings indicate that the pathogenic NY-1V G1 protein regulates cellular IFN responses upstream of IRF-3 phosphorylation at the level of the TBK-1 complex. These findings further suggest that the G1 cytoplasmic tail contains a virulence element which determines the ability of hantaviruses to bypass innate cellular immune responses and delineates a mechanism for pathogenic hantaviruses to successfully replicate within human endothelial cells.
Alff
Peter J
PJ
Molecular and Cellular Biology Graduate Program, SUNY at Stony Brook, Stony Brook, NY 11794, USA.
Gavrilovskaya
Irina N
IN
Gorbunova
Elena
E
Endriss
Karen
K
Chong
Yuson
Y
Geimonen
Erika
E
Sen
Nandini
N
Reich
Nancy C
NC
Mackow
Erich R
ER
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism*; D024882:Drug Resistance, Viral; D042783:Endothelial Cells / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics; D006476:Orthohantavirus / Q000145:classification / Q000472:pathogenicity* / Q000502:physiology; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000235:genetics; D007372:Interferons / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D014176:Protein Biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D020365:RNA Helicases / Q000378:metabolism*; D016415:Sequence Alignment; D014158:Transcription, Genetic / Q000235:genetics; D014779:Virus Replication
D050838:Interferon Regulatory Factor-3; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D007372:Interferons; D017346:Protein Serine-Threonine Kinases; C403191:TBK1 protein, human; D019204:GTP-Binding Proteins; D020365:RNA Helicases