title: Virulence factor NSs of rift valley fever virus recruits the F-box protein FBXO3 to degrade subunit p62 of general transcription factor TFIIH.
abstract: The nonstructural protein NSs is the main virulence factor of Rift Valley fever virus (RVFV; family Bunyaviridae, genus Phlebovirus), a serious pathogen of livestock and humans in Africa. RVFV NSs blocks transcriptional upregulation of antiviral type I interferons (IFN) and destroys the general transcription factor TFIIH subunit p62 via the ubiquitin/proteasome pathway. Here, we identified a subunit of E3 ubiquitin ligases, F-box protein FBXO3, as a host cell interactor of NSs. Small interfering RNA (siRNA)-mediated depletion of FBXO3 rescued p62 protein levels in RVFV-infected cells and elevated IFN transcription by 1 order of magnitude. NSs interacts with the full-length FBXO3 protein as well as with a truncated isoform that lacks the C-terminal acidic and poly(R)-rich domains. These isoforms are present in both the nucleus and the cytoplasm. NSs exclusively removes the nuclear pool of full-length FBXO3, likely due to consumption during the degradation process. F-box proteins form the variable substrate recognition subunit of the so-called SCF ubiquitin ligases, which also contain the constant components Skp1, cullin 1 (or cullin 7), and Rbx1. siRNA knockdown of Skp1 also protected p62 from degradation, suggesting involvement in NSs action. However, knockdown of cullin 1, cullin 7, or Rbx1 could not rescue p62 degradation by NSs. Our data show that the enzymatic removal of p62 via the host cell factor FBXO3 is a major mechanism of IFN suppression by RVFV.
  
  
  OBJECTIVE
  Rift Valley fever virus is a serious emerging pathogen of animals and humans. Its main virulence factor, NSs, enables unhindered virus replication by suppressing the antiviral innate immune system. We identified the E3 ubiquitin ligase FBXO3 as a novel host cell interactor of NSs. NSs recruits FBXO3 to destroy the general host cell transcription factor TFIIH-p62, resulting in suppression of the transcriptional upregulation of innate immunity.
authors.affiliation: Institute for Virology, Philipps-University Marburg, Marburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Kainulainen
authors.forename: Matthias
authors.initials: M
authors.lastname: Habjan
authors.forename: Philipp
authors.initials: P
authors.lastname: Hubel
authors.forename: Laura
authors.initials: L
authors.lastname: Busch
authors.forename: Simone
authors.initials: S
authors.lastname: Lau
authors.forename: Jacques
authors.initials: J
authors.lastname: Colinge
authors.forename: Giulio
authors.initials: G
authors.lastname: Superti-Furga
authors.forename: Andreas
authors.initials: A
authors.lastname: Pichlmair
authors.forename: Friedemann
authors.initials: F
authors.lastname: Weber
mesh_terms: D002460:Cell Line; D044783:F-Box Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D059748:Proteolysis; D012295:Rift Valley Fever / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D012296:Rift Valley fever virus / Q000235:genetics / Q000378:metabolism*; D051758:Transcription Factor TFIIH; D019473:Transcription Factors, TFII / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D044783:F-Box Proteins; D010750:Phosphoproteins; D051758:Transcription Factor TFIIH; D019473:Transcription Factors, TFII; D044767:Ubiquitin-Protein Ligases; D017361:Viral Nonstructural Proteins; D037521:Virulence Factors; C533903:FBXO3 protein, human; C072546:GTF2H1 protein, human