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Title: Protein Kinase R Degradation Is Essential for Rift Valley Fever Virus Infection and Is Regulated by SKP1-CUL1-F-box (SCF)FBXW11-NSs E3 Ligase

Abstract

Activated protein kinase R (PKR) plays a vital role in antiviral defense primarily by inhibiting protein synthesis and augmenting interferon responses. Many viral proteins have adopted unique strategies to counteract the deleterious effects of PKR. The NSs (Non-structural s) protein which is encoded by Rift Valley fever virus (RVFV) promotes early PKR proteasomal degradation through a previously undefined mechanism. In this study, we demonstrate that NSs carries out this activity by assembling the SCF (SKP1-CUL1-F-box)FBXW11 E3 ligase. NSs binds to the F-box protein, FBXW11, via the six amino acid sequence DDGFVE called the degron sequence and recruits PKR through an alternate binding site to the SCFFBXW11 E3 ligase. We further show that disrupting the assembly of the SCFFBXW11-NSs E3 ligase with MLN4924 (a small molecule inhibitor of SCF E3 ligase activity) or NSs degron viral mutants or siRNA knockdown of FBXW11 can block PKR degradation. Surprisingly, under these conditions when PKR degradation was blocked, NSs was essential and sufficient to activate PKR causing potent inhibition of RVFV infection by suppressing viral protein synthesis. These antiviral effects were antagonized by the loss of PKR expression or with a NSs deleted mutant virus. Therefore, early PKR activation by disassembly of SCFFBXW11-NSs E3more » ligase is sufficient to inhibit RVFV infection. Furthermore, FBXW11 and BTRC are the two homologues of the βTrCP (Beta-transducin repeat containing protein) gene that were previously described to be functionally redundant. However, in RVFV infection, among the two homologues of βTrCP, FBXW11 plays a dominant role in PKR degradation and is the limiting factor in the assembly of the SCFFBXW11 complex. Thus, FBXW11 serves as a master regulator of RVFV infection by promoting PKR degradation. Overall these findings provide new insights into NSs regulation of PKR activity and offer potential opportunities for therapeutic intervention of RVFV infection.« less

Authors:
 [1];  [1];  [1];  [2];  [1];  [1]
  1. United States Army Medical Research Inst. of Infectious Diseases, Frederick, MD (United States)
  2. Perkin Elmer, Waltham, MA (United States); United States Army Medical Research Inst. of Infectious Diseases, Frederick, MD (United States)
Publication Date:
Research Org.:
United States Army Medical Research Institute of Infectious Diseases, Frederick, Maryland (United States); Defense Threat Reduction Agency (DTRA), Joint Science and Technology Office for Chem Bio Defense (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1378555
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
PLoS Pathogens
Additional Journal Information:
Journal Volume: 12; Journal Issue: 2; Journal ID: ISSN 1553-7374
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Mudhasani, Rajini, Tran, Julie P., Retterer, Cary, Kota, Krishna P., Whitehouse, Chris A., and Bavari, Sina. Protein Kinase R Degradation Is Essential for Rift Valley Fever Virus Infection and Is Regulated by SKP1-CUL1-F-box (SCF)FBXW11-NSs E3 Ligase. United States: N. p., 2016. Web. doi:10.1371/journal.ppat.1005437.
Mudhasani, Rajini, Tran, Julie P., Retterer, Cary, Kota, Krishna P., Whitehouse, Chris A., & Bavari, Sina. Protein Kinase R Degradation Is Essential for Rift Valley Fever Virus Infection and Is Regulated by SKP1-CUL1-F-box (SCF)FBXW11-NSs E3 Ligase. United States. https://doi.org/10.1371/journal.ppat.1005437
Mudhasani, Rajini, Tran, Julie P., Retterer, Cary, Kota, Krishna P., Whitehouse, Chris A., and Bavari, Sina. 2016. "Protein Kinase R Degradation Is Essential for Rift Valley Fever Virus Infection and Is Regulated by SKP1-CUL1-F-box (SCF)FBXW11-NSs E3 Ligase". United States. https://doi.org/10.1371/journal.ppat.1005437. https://www.osti.gov/servlets/purl/1378555.
@article{osti_1378555,
title = {Protein Kinase R Degradation Is Essential for Rift Valley Fever Virus Infection and Is Regulated by SKP1-CUL1-F-box (SCF)FBXW11-NSs E3 Ligase},
author = {Mudhasani, Rajini and Tran, Julie P. and Retterer, Cary and Kota, Krishna P. and Whitehouse, Chris A. and Bavari, Sina},
abstractNote = {Activated protein kinase R (PKR) plays a vital role in antiviral defense primarily by inhibiting protein synthesis and augmenting interferon responses. Many viral proteins have adopted unique strategies to counteract the deleterious effects of PKR. The NSs (Non-structural s) protein which is encoded by Rift Valley fever virus (RVFV) promotes early PKR proteasomal degradation through a previously undefined mechanism. In this study, we demonstrate that NSs carries out this activity by assembling the SCF (SKP1-CUL1-F-box)FBXW11 E3 ligase. NSs binds to the F-box protein, FBXW11, via the six amino acid sequence DDGFVE called the degron sequence and recruits PKR through an alternate binding site to the SCFFBXW11 E3 ligase. We further show that disrupting the assembly of the SCFFBXW11-NSs E3 ligase with MLN4924 (a small molecule inhibitor of SCF E3 ligase activity) or NSs degron viral mutants or siRNA knockdown of FBXW11 can block PKR degradation. Surprisingly, under these conditions when PKR degradation was blocked, NSs was essential and sufficient to activate PKR causing potent inhibition of RVFV infection by suppressing viral protein synthesis. These antiviral effects were antagonized by the loss of PKR expression or with a NSs deleted mutant virus. Therefore, early PKR activation by disassembly of SCFFBXW11-NSs E3 ligase is sufficient to inhibit RVFV infection. Furthermore, FBXW11 and BTRC are the two homologues of the βTrCP (Beta-transducin repeat containing protein) gene that were previously described to be functionally redundant. However, in RVFV infection, among the two homologues of βTrCP, FBXW11 plays a dominant role in PKR degradation and is the limiting factor in the assembly of the SCFFBXW11 complex. Thus, FBXW11 serves as a master regulator of RVFV infection by promoting PKR degradation. Overall these findings provide new insights into NSs regulation of PKR activity and offer potential opportunities for therapeutic intervention of RVFV infection.},
doi = {10.1371/journal.ppat.1005437},
url = {https://www.osti.gov/biblio/1378555}, journal = {PLoS Pathogens},
issn = {1553-7374},
number = 2,
volume = 12,
place = {United States},
year = {Tue Feb 02 00:00:00 EST 2016},
month = {Tue Feb 02 00:00:00 EST 2016}
}

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Cited by: 42 works
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Works referenced in this record:

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The CUL1 C-Terminal Sequence and ROC1 Are Required for Efficient Nuclear Accumulation, NEDD8 Modification, and Ubiquitin Ligase Activity of CUL1
journal, January 2000


Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase
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WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 by DDB1-CUL4-ROC1 ligase
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NEDD8 Modification of CUL1 Dissociates p120CAND1, an Inhibitor of CUL1-SKP1 Binding and SCF Ligases
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NEDD8 Modification of CUL1 Dissociates p120CAND1, an Inhibitor of CUL1-SKP1 Binding and SCF Ligases
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Deregulated proteolysis by the F-box proteins SKP2 and β-TrCP: tipping the scales of cancer
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Function and regulation of cullin–RING ubiquitin ligases
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The many faces of β-TrCP E3 ubiquitin ligases: reflections in the magic mirror of cancer
journal, March 2004


Impaired degradation of inhibitory subunit of NF- B (I B) and  -catenin as a result of targeted disruption of the  -TrCP1 gene
journal, July 2003


A ΩX a V motif in the Rift Valley fever virus NSs protein is essential for degrading p62, forming nuclear filaments and virulence
journal, April 2015


A Dominant-negative UBC12 Mutant Sequesters NEDD8 and Inhibits NEDD8 Conjugation in Vivo
journal, May 2000


Induction of DNA Damage Signaling upon Rift Valley Fever Virus Infection Results in Cell Cycle Arrest and Increased Viral Replication
journal, January 2012


Tipping the Balance: Antagonism of PKR Kinase and ADAR1 Deaminase Functions by Virus Gene Products
journal, September 2009


Systematic analysis and nomenclature of mammalian F-box proteins
journal, November 2004


The CUL1 C-Terminal Sequence and ROC1 Are Required for Efficient Nuclear Accumulation, NEDD8 Modification, and Ubiquitin Ligase Activity of CUL1
journal, January 2000


NSs Protein of Rift Valley Fever Virus Induces the Specific Degradation of the Double-Stranded RNA-Dependent Protein Kinase
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Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase
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High-Content Image–Based Screening of a Signal Transduction Pathway Inhibitor Small-Molecule Library against Highly Pathogenic RNA Viruses
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Works referencing / citing this record:

Viral proteins targeting host protein kinase R to evade an innate immune response: a mini review
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NSs Virulence Factor of Rift Valley Fever Virus Engages the F-Box Proteins FBXW11 and β-TRCP1 To Degrade the Antiviral Protein Kinase PKR
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NSs Protein of Sandfly Fever Sicilian Phlebovirus Counteracts Interferon (IFN) Induction by Masking the DNA-Binding Domain of IFN Regulatory Factor 3
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Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells
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Toscana virus non-structural protein NSs acts as E3 ubiquitin ligase promoting RIG-I degradation
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Cellular Cullin RING Ubiquitin Ligases: Druggable Host Dependency Factors of Cytomegaloviruses
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Alterations in the host transcriptome in vitro following Rift Valley fever virus infection
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VHS, US3 and UL13 viral tegument proteins are required for Herpes Simplex Virus-Induced modification of protein kinase R
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NSs Virulence Factor of Rift Valley Fever Virus Engages the F-Box Proteins FBXW11 and β-TRCP1 To Degrade the Antiviral Protein Kinase PKR
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NSs Protein of Sandfly Fever Sicilian Phlebovirus Counteracts Interferon (IFN) Induction by Masking the DNA-Binding Domain of IFN Regulatory Factor 3
journal, September 2018


Mechanistic Insight into the Host Transcription Inhibition Function of Rift Valley Fever Virus NSs and Its Importance in Virulence
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Toscana virus non-structural protein NSs acts as E3 ubiquitin ligase promoting RIG-I degradation
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Cellular Cullin RING Ubiquitin Ligases: Druggable Host Dependency Factors of Cytomegaloviruses
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Phleboviruses and the Type I Interferon Response
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