Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease

Linda Wiltzer, Kazuma Okada, Satoko Yamaoka, Florence Larrous, Henna V Kuusisto, Makoto Sugiyama, Danielle Blondel, Herve Bourhy, David A Jans, Naoto Ito, Gregory W Moseley

Research output: Contribution to journalArticleResearchpeer-review

Abstract

BACKGROUND: Rabies virus (RABV) causes rabies disease resulting in >55,000 human deaths/year. The multifunctional RABV P-protein has essential roles in genome replication, and forms interactions with cellular STAT proteins that are thought to underlie viral antagonism of interferon-dependent immunity. However, the molecular details of P-protein-STAT interaction, and its importance to disease are unresolved. METHODS: Studies were performed using sequence/structure analysis, mutagenesis, immunoprecipitation, luciferase and qRT-PCR-based signaling assays, confocal microscopy and reverse genetics/in vivo infection. RESULTS: We identified a hydrophobic pocket of the P-protein C-terminal domain as critical to STAT-binding/antagonism. This interface was found to be functionally and spatially independent of the region responsible for N-protein interaction, which is critical to genome replication. Based on these findings, we generated the first mutant RABV lacking STAT-association. Growth of the virus in vitro was unimpaired, but it lacked STAT-antagonist function and was highly sensitive to interferon. Importantly, growth of the virus was strongly attenuated in brains of infected mice, producing no major neurological symptoms, compared with the invariably lethal wild-type virus. CONCLUSIONS: These data represent direct evidence that P-protein-STAT interaction is critical to rabies, and provide novel insights into the mechanism by which RABV coordinates distinct functions in interferon antagonism and replication.
Original languageEnglish
Pages (from-to)1744 - 1753
Number of pages10
JournalJournal of Infectious Diseases
Volume209
Issue number11
DOIs
Publication statusPublished - 2014

Cite this

Wiltzer, Linda ; Okada, Kazuma ; Yamaoka, Satoko ; Larrous, Florence ; Kuusisto, Henna V ; Sugiyama, Makoto ; Blondel, Danielle ; Bourhy, Herve ; Jans, David A ; Ito, Naoto ; Moseley, Gregory W. / Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease. In: Journal of Infectious Diseases. 2014 ; Vol. 209, No. 11. pp. 1744 - 1753.
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title = "Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease",
abstract = "BACKGROUND: Rabies virus (RABV) causes rabies disease resulting in >55,000 human deaths/year. The multifunctional RABV P-protein has essential roles in genome replication, and forms interactions with cellular STAT proteins that are thought to underlie viral antagonism of interferon-dependent immunity. However, the molecular details of P-protein-STAT interaction, and its importance to disease are unresolved. METHODS: Studies were performed using sequence/structure analysis, mutagenesis, immunoprecipitation, luciferase and qRT-PCR-based signaling assays, confocal microscopy and reverse genetics/in vivo infection. RESULTS: We identified a hydrophobic pocket of the P-protein C-terminal domain as critical to STAT-binding/antagonism. This interface was found to be functionally and spatially independent of the region responsible for N-protein interaction, which is critical to genome replication. Based on these findings, we generated the first mutant RABV lacking STAT-association. Growth of the virus in vitro was unimpaired, but it lacked STAT-antagonist function and was highly sensitive to interferon. Importantly, growth of the virus was strongly attenuated in brains of infected mice, producing no major neurological symptoms, compared with the invariably lethal wild-type virus. CONCLUSIONS: These data represent direct evidence that P-protein-STAT interaction is critical to rabies, and provide novel insights into the mechanism by which RABV coordinates distinct functions in interferon antagonism and replication.",
author = "Linda Wiltzer and Kazuma Okada and Satoko Yamaoka and Florence Larrous and Kuusisto, {Henna V} and Makoto Sugiyama and Danielle Blondel and Herve Bourhy and Jans, {David A} and Naoto Ito and Moseley, {Gregory W}",
year = "2014",
doi = "10.1093/infdis/jit829",
language = "English",
volume = "209",
pages = "1744 -- 1753",
journal = "Journal of Infectious Diseases",
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Wiltzer, L, Okada, K, Yamaoka, S, Larrous, F, Kuusisto, HV, Sugiyama, M, Blondel, D, Bourhy, H, Jans, DA, Ito, N & Moseley, GW 2014, 'Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease' Journal of Infectious Diseases, vol. 209, no. 11, pp. 1744 - 1753. https://doi.org/10.1093/infdis/jit829

Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease. / Wiltzer, Linda; Okada, Kazuma; Yamaoka, Satoko; Larrous, Florence; Kuusisto, Henna V; Sugiyama, Makoto; Blondel, Danielle; Bourhy, Herve; Jans, David A; Ito, Naoto; Moseley, Gregory W.

In: Journal of Infectious Diseases, Vol. 209, No. 11, 2014, p. 1744 - 1753.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease

AU - Wiltzer, Linda

AU - Okada, Kazuma

AU - Yamaoka, Satoko

AU - Larrous, Florence

AU - Kuusisto, Henna V

AU - Sugiyama, Makoto

AU - Blondel, Danielle

AU - Bourhy, Herve

AU - Jans, David A

AU - Ito, Naoto

AU - Moseley, Gregory W

PY - 2014

Y1 - 2014

N2 - BACKGROUND: Rabies virus (RABV) causes rabies disease resulting in >55,000 human deaths/year. The multifunctional RABV P-protein has essential roles in genome replication, and forms interactions with cellular STAT proteins that are thought to underlie viral antagonism of interferon-dependent immunity. However, the molecular details of P-protein-STAT interaction, and its importance to disease are unresolved. METHODS: Studies were performed using sequence/structure analysis, mutagenesis, immunoprecipitation, luciferase and qRT-PCR-based signaling assays, confocal microscopy and reverse genetics/in vivo infection. RESULTS: We identified a hydrophobic pocket of the P-protein C-terminal domain as critical to STAT-binding/antagonism. This interface was found to be functionally and spatially independent of the region responsible for N-protein interaction, which is critical to genome replication. Based on these findings, we generated the first mutant RABV lacking STAT-association. Growth of the virus in vitro was unimpaired, but it lacked STAT-antagonist function and was highly sensitive to interferon. Importantly, growth of the virus was strongly attenuated in brains of infected mice, producing no major neurological symptoms, compared with the invariably lethal wild-type virus. CONCLUSIONS: These data represent direct evidence that P-protein-STAT interaction is critical to rabies, and provide novel insights into the mechanism by which RABV coordinates distinct functions in interferon antagonism and replication.

AB - BACKGROUND: Rabies virus (RABV) causes rabies disease resulting in >55,000 human deaths/year. The multifunctional RABV P-protein has essential roles in genome replication, and forms interactions with cellular STAT proteins that are thought to underlie viral antagonism of interferon-dependent immunity. However, the molecular details of P-protein-STAT interaction, and its importance to disease are unresolved. METHODS: Studies were performed using sequence/structure analysis, mutagenesis, immunoprecipitation, luciferase and qRT-PCR-based signaling assays, confocal microscopy and reverse genetics/in vivo infection. RESULTS: We identified a hydrophobic pocket of the P-protein C-terminal domain as critical to STAT-binding/antagonism. This interface was found to be functionally and spatially independent of the region responsible for N-protein interaction, which is critical to genome replication. Based on these findings, we generated the first mutant RABV lacking STAT-association. Growth of the virus in vitro was unimpaired, but it lacked STAT-antagonist function and was highly sensitive to interferon. Importantly, growth of the virus was strongly attenuated in brains of infected mice, producing no major neurological symptoms, compared with the invariably lethal wild-type virus. CONCLUSIONS: These data represent direct evidence that P-protein-STAT interaction is critical to rabies, and provide novel insights into the mechanism by which RABV coordinates distinct functions in interferon antagonism and replication.

UR - http://jid.oxfordjournals.org/content/209/11/1744.full.pdf+html

U2 - 10.1093/infdis/jit829

DO - 10.1093/infdis/jit829

M3 - Article

VL - 209

SP - 1744

EP - 1753

JO - Journal of Infectious Diseases

JF - Journal of Infectious Diseases

SN - 0022-1899

IS - 11

ER -