Disruption of tlr3 signaling due to cleavage of trif by the hepatitis a virus protease-polymerase processing intermediate, 3cd

Lin Qu, Zongdi Feng, Daisuke Yamane, Yuqiong Liang, Robert E. Lanford, Kui Li, Stanley M. Lemon

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Toll-like receptor 3 (TLR3) and cytosolic RIG-I-like helicases (RIG-I and MDA5) sense viral RNAs and activate innate immune signaling pathways that induce expression of interferon (IFN) through specific adaptor proteins, TIR domain-containing adaptor inducing interferon-β (TRIF), and mitochondrial antiviral signaling protein (MAVS), respectively. Previously, we demonstrated that hepatitis A virus (HAV), a unique hepatotropic human picornavirus, disrupts RIG-I/MDA5 signaling by targeting MAVS for cleavage by 3ABC, a precursor of the sole HAV protease, 3C pro, that is derived by auto-processing of the P3 (3ABCD) segment of the viral polyprotein. Here, we show that HAV also disrupts TLR3 signaling, inhibiting poly(I:C)-stimulated dimerization of IFN regulatory factor 3 (IRF-3), IRF-3 translocation to the nucleus, and IFN-β promoter activation, by targeting TRIF for degradation by a distinct 3ABCD processing intermediate, the 3CD protease-polymerase precursor. TRIF is proteolytically cleaved by 3CD, but not by the mature 3C pro protease or the 3ABC precursor that degrades MAVS. 3CD-mediated degradation of TRIF depends on both the cysteine protease activity of 3C pro and downstream 3D pol sequence, but not 3D pol polymerase activity. Cleavage occurs at two non-canonical 3C pro recognition sequences in TRIF, and involves a hierarchical process in which primary cleavage at Gln-554 is a prerequisite for scission at Gln-190. The results of mutational studies indicate that 3D pol sequence modulates the substrate specificity of the upstream 3C pro protease when fused to it in cis in 3CD, allowing 3CD to target cleavage sites not normally recognized by 3C pro. HAV thus disrupts both RIG-I/MDA5 and TLR3 signaling pathways through cleavage of essential adaptor proteins by two distinct protease precursors derived from the common 3ABCD polyprotein processing intermediate.

Original languageEnglish (US)
Article numbere1002169
JournalPLoS Pathogens
Volume7
Issue number9
DOIs
StatePublished - Sep 1 2011

Fingerprint

Hepatitis Viruses
Toll-Like Receptor 3
Hepatitis A virus
Peptide Hydrolases
Interferon Regulatory Factor-3
Interferons
Antiviral Agents
Polyproteins
Proteins
Picornaviridae
Cysteine Proteases
Viral RNA
Dimerization
Substrate Specificity
3C proteases

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Microbiology
  • Immunology
  • Molecular Biology
  • Genetics
  • Virology

Cite this

Disruption of tlr3 signaling due to cleavage of trif by the hepatitis a virus protease-polymerase processing intermediate, 3cd. / Qu, Lin; Feng, Zongdi; Yamane, Daisuke; Liang, Yuqiong; Lanford, Robert E.; Li, Kui; Lemon, Stanley M.

In: PLoS Pathogens, Vol. 7, No. 9, e1002169, 01.09.2011.

Research output: Contribution to journalArticle

Qu, Lin ; Feng, Zongdi ; Yamane, Daisuke ; Liang, Yuqiong ; Lanford, Robert E. ; Li, Kui ; Lemon, Stanley M. / Disruption of tlr3 signaling due to cleavage of trif by the hepatitis a virus protease-polymerase processing intermediate, 3cd. In: PLoS Pathogens. 2011 ; Vol. 7, No. 9.
@article{f0c1064cccec426f8f6b2979d8c8c7b3,
title = "Disruption of tlr3 signaling due to cleavage of trif by the hepatitis a virus protease-polymerase processing intermediate, 3cd",
abstract = "Toll-like receptor 3 (TLR3) and cytosolic RIG-I-like helicases (RIG-I and MDA5) sense viral RNAs and activate innate immune signaling pathways that induce expression of interferon (IFN) through specific adaptor proteins, TIR domain-containing adaptor inducing interferon-β (TRIF), and mitochondrial antiviral signaling protein (MAVS), respectively. Previously, we demonstrated that hepatitis A virus (HAV), a unique hepatotropic human picornavirus, disrupts RIG-I/MDA5 signaling by targeting MAVS for cleavage by 3ABC, a precursor of the sole HAV protease, 3C pro, that is derived by auto-processing of the P3 (3ABCD) segment of the viral polyprotein. Here, we show that HAV also disrupts TLR3 signaling, inhibiting poly(I:C)-stimulated dimerization of IFN regulatory factor 3 (IRF-3), IRF-3 translocation to the nucleus, and IFN-β promoter activation, by targeting TRIF for degradation by a distinct 3ABCD processing intermediate, the 3CD protease-polymerase precursor. TRIF is proteolytically cleaved by 3CD, but not by the mature 3C pro protease or the 3ABC precursor that degrades MAVS. 3CD-mediated degradation of TRIF depends on both the cysteine protease activity of 3C pro and downstream 3D pol sequence, but not 3D pol polymerase activity. Cleavage occurs at two non-canonical 3C pro recognition sequences in TRIF, and involves a hierarchical process in which primary cleavage at Gln-554 is a prerequisite for scission at Gln-190. The results of mutational studies indicate that 3D pol sequence modulates the substrate specificity of the upstream 3C pro protease when fused to it in cis in 3CD, allowing 3CD to target cleavage sites not normally recognized by 3C pro. HAV thus disrupts both RIG-I/MDA5 and TLR3 signaling pathways through cleavage of essential adaptor proteins by two distinct protease precursors derived from the common 3ABCD polyprotein processing intermediate.",
author = "Lin Qu and Zongdi Feng and Daisuke Yamane and Yuqiong Liang and Lanford, {Robert E.} and Kui Li and Lemon, {Stanley M.}",
year = "2011",
month = "9",
day = "1",
doi = "10.1371/journal.ppat.1002169",
language = "English (US)",
volume = "7",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "9",

}

TY - JOUR

T1 - Disruption of tlr3 signaling due to cleavage of trif by the hepatitis a virus protease-polymerase processing intermediate, 3cd

AU - Qu, Lin

AU - Feng, Zongdi

AU - Yamane, Daisuke

AU - Liang, Yuqiong

AU - Lanford, Robert E.

AU - Li, Kui

AU - Lemon, Stanley M.

PY - 2011/9/1

Y1 - 2011/9/1

N2 - Toll-like receptor 3 (TLR3) and cytosolic RIG-I-like helicases (RIG-I and MDA5) sense viral RNAs and activate innate immune signaling pathways that induce expression of interferon (IFN) through specific adaptor proteins, TIR domain-containing adaptor inducing interferon-β (TRIF), and mitochondrial antiviral signaling protein (MAVS), respectively. Previously, we demonstrated that hepatitis A virus (HAV), a unique hepatotropic human picornavirus, disrupts RIG-I/MDA5 signaling by targeting MAVS for cleavage by 3ABC, a precursor of the sole HAV protease, 3C pro, that is derived by auto-processing of the P3 (3ABCD) segment of the viral polyprotein. Here, we show that HAV also disrupts TLR3 signaling, inhibiting poly(I:C)-stimulated dimerization of IFN regulatory factor 3 (IRF-3), IRF-3 translocation to the nucleus, and IFN-β promoter activation, by targeting TRIF for degradation by a distinct 3ABCD processing intermediate, the 3CD protease-polymerase precursor. TRIF is proteolytically cleaved by 3CD, but not by the mature 3C pro protease or the 3ABC precursor that degrades MAVS. 3CD-mediated degradation of TRIF depends on both the cysteine protease activity of 3C pro and downstream 3D pol sequence, but not 3D pol polymerase activity. Cleavage occurs at two non-canonical 3C pro recognition sequences in TRIF, and involves a hierarchical process in which primary cleavage at Gln-554 is a prerequisite for scission at Gln-190. The results of mutational studies indicate that 3D pol sequence modulates the substrate specificity of the upstream 3C pro protease when fused to it in cis in 3CD, allowing 3CD to target cleavage sites not normally recognized by 3C pro. HAV thus disrupts both RIG-I/MDA5 and TLR3 signaling pathways through cleavage of essential adaptor proteins by two distinct protease precursors derived from the common 3ABCD polyprotein processing intermediate.

AB - Toll-like receptor 3 (TLR3) and cytosolic RIG-I-like helicases (RIG-I and MDA5) sense viral RNAs and activate innate immune signaling pathways that induce expression of interferon (IFN) through specific adaptor proteins, TIR domain-containing adaptor inducing interferon-β (TRIF), and mitochondrial antiviral signaling protein (MAVS), respectively. Previously, we demonstrated that hepatitis A virus (HAV), a unique hepatotropic human picornavirus, disrupts RIG-I/MDA5 signaling by targeting MAVS for cleavage by 3ABC, a precursor of the sole HAV protease, 3C pro, that is derived by auto-processing of the P3 (3ABCD) segment of the viral polyprotein. Here, we show that HAV also disrupts TLR3 signaling, inhibiting poly(I:C)-stimulated dimerization of IFN regulatory factor 3 (IRF-3), IRF-3 translocation to the nucleus, and IFN-β promoter activation, by targeting TRIF for degradation by a distinct 3ABCD processing intermediate, the 3CD protease-polymerase precursor. TRIF is proteolytically cleaved by 3CD, but not by the mature 3C pro protease or the 3ABC precursor that degrades MAVS. 3CD-mediated degradation of TRIF depends on both the cysteine protease activity of 3C pro and downstream 3D pol sequence, but not 3D pol polymerase activity. Cleavage occurs at two non-canonical 3C pro recognition sequences in TRIF, and involves a hierarchical process in which primary cleavage at Gln-554 is a prerequisite for scission at Gln-190. The results of mutational studies indicate that 3D pol sequence modulates the substrate specificity of the upstream 3C pro protease when fused to it in cis in 3CD, allowing 3CD to target cleavage sites not normally recognized by 3C pro. HAV thus disrupts both RIG-I/MDA5 and TLR3 signaling pathways through cleavage of essential adaptor proteins by two distinct protease precursors derived from the common 3ABCD polyprotein processing intermediate.

UR - http://www.scopus.com/inward/record.url?scp=79960598740&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79960598740&partnerID=8YFLogxK

U2 - 10.1371/journal.ppat.1002169

DO - 10.1371/journal.ppat.1002169

M3 - Article

VL - 7

JO - PLoS Pathogens

JF - PLoS Pathogens

SN - 1553-7366

IS - 9

M1 - e1002169

ER -