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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

Journal Article · · PLoS Pathogens
 [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)
+ Show Author Affiliations
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.
Research Organization:
Defense Threat Reduction Agency (DTRA), Joint Science and Technology Office for Chem Bio Defense (United States); United States Army Medical Research Institute of Infectious Diseases, Frederick, Maryland (United States)
Sponsoring Organization:
USDOE
OSTI ID:
1378555
Journal Information:
PLoS Pathogens, Journal Name: PLoS Pathogens Journal Issue: 2 Vol. 12; ISSN 1553-7374
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English

References (68)

Specificity, location and function of βTrCP isoforms and their splice variants journal April 2011
DEPTOR, an mTOR Inhibitor, Is a Physiological Substrate of SCFβTrCP E3 Ubiquitin Ligase and Regulates Survival and Autophagy journal October 2011
Evaluation of the efficacy and safety of the Rift Valley Fever Clone 13 vaccine in sheep journal June 2010
The cytoplasmic tail of hantavirus Gn glycoprotein interacts with RNA journal September 2011
Rift Valley fever virus NSs inhibits host transcription independently of the degradation of dsRNA-dependent protein kinase PKR journal January 2013
Role of the cytosolic tails of Rift Valley fever virus envelope glycoproteins in viral morphogenesis journal January 2014
ROC1, a Homolog of APC11, Represents a Family of Cullin Partners with an Associated Ubiquitin Ligase Activity journal April 1999
NEDD8 Modification of CUL1 Dissociates p120CAND1, an Inhibitor of CUL1-SKP1 Binding and SCF Ligases journal December 2002
Control of Meiotic and Mitotic Progression by the F Box Protein β-Trcp1 In Vivo journal June 2003
An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer journal April 2009
Deregulated proteolysis by the F-box proteins SKP2 and β-TrCP: tipping the scales of cancer journal June 2008
Function and regulation of cullin–RING ubiquitin ligases journal January 2005
The many faces of β-TrCP E3 ubiquitin ligases: reflections in the magic mirror of cancer journal March 2004
Cytoplasmic localized ubiquitin ligase cullin 7 binds to p53 and promotes cell growth by antagonizing p53 function journal March 2006
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
Rapid Detection and Quantification of RNA of Ebola and Marburg Viruses, Lassa Virus, Crimean-Congo Hemorrhagic Fever Virus, Rift Valley Fever Virus, Dengue Virus, and Yellow Fever Virus by Real-Time Reverse Transcription-PCR journal July 2002
Role of the Cytoplasmic Tail Domains of Bunyamwera Orthobunyavirus Glycoproteins Gn and Gc in Virus Assembly and Morphogenesis journal July 2007
Loss of Protein Kinase PKR Expression in Human HeLa Cells Complements the Vaccinia Virus E3L Deletion Mutant Phenotype by Restoration of Viral Protein Synthesis journal October 2007
NSs Protein of Rift Valley Fever Virus Induces the Specific Degradation of the Double-Stranded RNA-Dependent Protein Kinase journal February 2009
NSs Protein of Rift Valley Fever Virus Promotes Posttranslational Downregulation of the TFIIH Subunit p62 journal May 2011
Virulence Factor NSs of Rift Valley Fever Virus Recruits the F-Box Protein FBXO3 To Degrade Subunit p62 of General Transcription Factor TFIIH journal January 2014
IFITM-2 and IFITM-3 but Not IFITM-1 Restrict Rift Valley Fever Virus journal May 2013
The cellular 68,000-Mr protein kinase is highly autophosphorylated and activated yet significantly degraded during poliovirus infection: implications for translational regulation. journal January 1989
Genetic Evidence for an Interferon-Antagonistic Function of Rift Valley Fever Virus Nonstructural Protein NSs journal February 2001
Rescue of Infectious Rift Valley Fever Virus Entirely from cDNA, Analysis of Virus Lacking the NSs Gene, and Expression of a Foreign Gene journal February 2006
Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase journal October 2014
High-Content Image–Based Screening of a Signal Transduction Pathway Inhibitor Small-Molecule Library against Highly Pathogenic RNA Viruses journal January 2015
Transcriptome and de novo analysis of Rosa xanthina f. spontanea in response to cold stress journal October 2021
Rapid Accumulation of Virulent Rift Valley Fever Virus in Mice from an Attenuated Virus Carrying a Single Nucleotide Substitution in the M RNA journal April 2010
Rift Valley Fever Virus NSs Protein Promotes Post-Transcriptional Downregulation of Protein Kinase PKR and Inhibits eIF2α Phosphorylation journal February 2009
Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method journal December 2001
Identification of Substrates for F‐Box Proteins book December 2005
ROC1, a Homolog of APC11, Represents a Family of Cullin Partners with an Associated Ubiquitin Ligase Activity journal April 1999
NEDD8 Modification of CUL1 Dissociates p120CAND1, an Inhibitor of CUL1-SKP1 Binding and SCF Ligases journal December 2002
Control of Meiotic and Mitotic Progression by the F Box Protein β-Trcp1 In Vivo journal June 2003
Reactive oxygen species activate NFκB (p65) and p53 and induce apoptosis in RVFV infected liver cells journal January 2014
Nedd8 on cullin: building an expressway to protein destruction journal March 2004
Critical role for IL-1β in DNA damage-induced mucositis journal January 2014
Homodimer of Two F-box Proteins βTrCP1 or βTrCP2 Binds to IκBα for Signal-dependent Ubiquitination journal January 2000
Induction of DNA Damage Signaling upon Rift Valley Fever Virus Infection Results in Cell Cycle Arrest and Increased Viral Replication journal January 2012
WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 by DDB1-CUL4-ROC1 ligase journal April 2008
Rapid Detection and Quantification of RNA of Ebola and Marburg Viruses, Lassa Virus, Crimean-Congo Hemorrhagic Fever Virus, Rift Valley Fever Virus, Dengue Virus, and Yellow Fever Virus by Real-Time Reverse Transcription-PCR journal July 2002
Role of the Cytoplasmic Tail Domains of Bunyamwera Orthobunyavirus Glycoproteins Gn and Gc in Virus Assembly and Morphogenesis journal July 2007
Loss of Protein Kinase PKR Expression in Human HeLa Cells Complements the Vaccinia Virus E3L Deletion Mutant Phenotype by Restoration of Viral Protein Synthesis journal October 2007
Acid-Activated Structural Reorganization of the Rift Valley Fever Virus Gc Fusion Protein journal October 2012
NSs Protein of Rift Valley Fever Virus Induces the Specific Degradation of the Double-Stranded RNA-Dependent Protein Kinase journal February 2009
NSs Protein of Rift Valley Fever Virus Promotes Posttranslational Downregulation of the TFIIH Subunit p62 journal May 2011
Virulence Factor NSs of Rift Valley Fever Virus Recruits the F-Box Protein FBXO3 To Degrade Subunit p62 of General Transcription Factor TFIIH journal January 2014
IFITM-2 and IFITM-3 but Not IFITM-1 Restrict Rift Valley Fever Virus journal May 2013
The cellular 68,000-Mr protein kinase is highly autophosphorylated and activated yet significantly degraded during poliovirus infection: implications for translational regulation. journal January 1989
Genetic Evidence for an Interferon-Antagonistic Function of Rift Valley Fever Virus Nonstructural Protein NSs journal February 2001
Double-Stranded RNA Is Produced by Positive-Strand RNA Viruses and DNA Viruses but Not in Detectable Amounts by Negative-Strand RNA Viruses journal May 2006
Rescue of Infectious Rift Valley Fever Virus Entirely from cDNA, Analysis of Virus Lacking the NSs Gene, and Expression of a Foreign Gene journal February 2006
Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase journal October 2014
The CUL1 C-Terminal Sequence and ROC1 Are Required for Efficient Nuclear Accumulation, NEDD8 Modification, and Ubiquitin Ligase Activity of CUL1 journal November 2000
Development of High-Content Imaging Assays for Lethal Viral Pathogens journal August 2010
High Content Image-Based Screening of a Protease Inhibitor Library Reveals Compounds Broadly Active against Rift Valley Fever Virus and Other Highly Pathogenic RNA Viruses journal August 2014
Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase text January 2015
WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 by DDB1-CUL4-ROC1 ligase text January 2008
NEDD8 Modification of CUL1 Dissociates p120CAND1, an Inhibitor of CUL1-SKP1 Binding and SCF Ligases text January 2002
ROC1, a Homolog of APC11, Represents a Family of Cullin Partners with an Associated Ubiquitin Ligase Activity text January 1999
Characterization of Clone 13, a Naturally Attenuated Avirulent Isolate of Rift Valley Fever Virus, which is Altered in the Small Segment * journal October 1995

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Alterations in the host transcriptome in vitro following Rift Valley fever virus infection journal October 2017
A single-cycle replicable Rift Valley fever phlebovirus vaccine carrying a mutated NSs confers full protection from lethal challenge in mice journal November 2018
VHS, US3 and UL13 viral tegument proteins are required for Herpes Simplex Virus-Induced modification of protein kinase R journal March 2020
Enhanced replication of mouse adenovirus type 1 following virus-induced degradation of protein kinase R (PKR) journal March 2019
NSs Virulence Factor of Rift Valley Fever Virus Engages the F-Box Proteins FBXW11 and β-TRCP1 To Degrade the Antiviral Protein Kinase PKR journal April 2016
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 journal October 2016
Toscana virus non-structural protein NSs acts as E3 ubiquitin ligase promoting RIG-I degradation journal December 2019
Cellular Cullin RING Ubiquitin Ligases: Druggable Host Dependency Factors of Cytomegaloviruses journal April 2019
Phleboviruses and the Type I Interferon Response journal June 2016
Characterization of the non-structural (NSs) protein of tick-borne phleboviruses text January 2017
Viral proteins targeting host protein kinase R to evade an innate immune response: a mini review journal January 2018
Translational Control in Virus-Infected Cells journal June 2018
Rift Valley fever virus NSs protein functions and the similarity to other bunyavirus NSs proteins journal July 2016
Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells journal November 2019

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