Molecular determinants of TRIF proteolysis mediated by the hepatitis C virus NS3/4A protease

Josephine C. Ferreon, Allan Chris M. Ferreon, Kui Li, Stanley M. Lemon

Research output: Contribution to journalArticle

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Abstract

Persistent infections with hepatitis C virus (HCV) are a major cause of liver disease and reflect its ability to disrupt virus-induced signaling pathways activating cellular antiviral defenses. HCV evasion of double-stranded RNA signaling through Toll-like receptor 3 is mediated by the viral protease NS3/4A, which directs proteolysis of its proline-rich adaptor protein, Toll-IL-1 receptor domain containing adaptor-inducing interferon-β (TRIF). The TRIF cleavage site has remarkable homology with the viral NS4B/5A substrate, although an 8-residue polyproline track extends upstream from the P6 position in lieu of the acidic residue present in viral substrates. Circular dichroism (CD) spectroscopy confirmed that a substantial fraction of TRIF exists as polyproline II helices, and inclusion of the polyproline track increased affinity of P side TRIF peptides for the HCV-BK protease. A polyproline II peptide representing an SH3 binding motif (PPPVPPRRR, Sos) bound NS3 with moderate affinity, resulting in inhibition of proteolytic activity. Chemical shift perturbations in NMR spectra indicated that Sos binds a 310 helix close to the protease active site. Thus, a polyproline II interaction with the 310 helix likely facilitates NS3/4A recognition of TRIF, indicating a significant difference from NS3/4A recognition of viral substrates. Because SH3 binding motifs are also present in NS5A, a viral protein that interacts with NS3, we speculate that the NS3 310 helix may be a site of interaction with other viral proteins.

Original languageEnglish (US)
Pages (from-to)20483-20492
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number21
DOIs
StatePublished - May 27 2005
Externally publishedYes

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Proteolysis
Viruses
Peptide Hydrolases
Hepacivirus
Viral Proteins
Substrates
Toll-Like Receptor 3
Circular dichroism spectroscopy
Peptides
Interleukin-1 Receptors
Double-Stranded RNA
Chemical shift
Circular Dichroism
Proline
Liver
Interferons
Antiviral Agents
Liver Diseases
Catalytic Domain
Spectrum Analysis

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Molecular determinants of TRIF proteolysis mediated by the hepatitis C virus NS3/4A protease. / Ferreon, Josephine C.; Ferreon, Allan Chris M.; Li, Kui; Lemon, Stanley M.

In: Journal of Biological Chemistry, Vol. 280, No. 21, 27.05.2005, p. 20483-20492.

Research output: Contribution to journalArticle

Ferreon, Josephine C. ; Ferreon, Allan Chris M. ; Li, Kui ; Lemon, Stanley M. / Molecular determinants of TRIF proteolysis mediated by the hepatitis C virus NS3/4A protease. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 21. pp. 20483-20492.
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