PMID- 30944180
OWN - NLM
STAT- MEDLINE
DCOM- 20200623
LR  - 20200623
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 93
IP  - 12
DP  - 2019 Jun 15
TI  - Arterivirus nsp4 Antagonizes Interferon Beta Production by Proteolytically 
      Cleaving NEMO at Multiple Sites.
LID - 10.1128/JVI.00385-19 [doi]
LID - e00385-19
AB  - Equine arteritis virus (EAV) and porcine reproductive and respiratory syndrome 
      virus (PRRSV) represent two members of the family Arteriviridae and pose major 
      threats for the horse- and swine-breeding industries worldwide. A previous study 
      suggested that PRRSV nsp4, a 3C-like protease, antagonizes interferon beta 
      (IFN-beta) production by cleaving the NF-kappaB essential modulator (NEMO) at a single 
      site, glutamate 349 (E349). Here, we demonstrated that EAV nsp4 also inhibited 
      virus-induced IFN-beta production by targeting NEMO for proteolytic cleavage and 
      that the scission occurred at four sites: E166, E171, glutamine 205 (Q205), and 
      E349. Additionally, we found that, besides the previously reported cleavage site 
      E349 in NEMO, scission by PRRSV nsp4 took place at two additional sites, E166 and 
      E171. These results imply that while cleaving NEMO is a common strategy utilized 
      by EAV and PRRSV nsp4 to antagonize IFN induction, EAV nsp4 adopts a more complex 
      substrate recognition mechanism to target NEMO. By analyzing the abilities of the 
      eight different NEMO fragments resulting from EAV or PRRSV nsp4 scission to 
      induce IFN-beta production, we serendipitously found that a NEMO fragment (residues 
      1 to 349) could activate IFN-beta transcription more robustly than full-length NEMO, 
      whereas all other NEMO cleavage products were abrogated for the IFN-beta-inducing 
      capacity. Thus, NEMO cleavage at E349 alone may not be sufficient to completely 
      inactivate the IFN response via this signaling adaptor. Altogether, our findings 
      suggest that EAV and PRRSV nsp4 cleave NEMO at multiple sites and that this 
      strategy is critical for disarming the innate immune response for viral 
      survival.IMPORTANCE The arterivirus nsp4-encoded 3C-like protease (3CL(pro)) 
      plays an important role in virus replication and immune evasion, making it an 
      attractive target for antiviral therapeutics. Previous work suggested that PRRSV 
      nsp4 suppresses type I IFN production by cleaving NEMO at a single site. In 
      contrast, the present study demonstrates that both EAV and PRRSV nsp4 cleave NEMO 
      at multiple sites and that this strategy is essential for disruption of type I 
      IFN production. Moreover, we reveal that EAV nsp4 also cleaves NEMO at glutamine 
      205 (Q205), which is not targeted by PRRSV nsp4. Notably, targeting a glutamine 
      in NEMO for cleavage has been observed only with picornavirus 3C proteases 
      (3C(pro)) and coronavirus 3CL(pro) In aggregate, our work expands knowledge of 
      the innate immune evasion mechanisms associated with NEMO cleavage by arterivirus 
      nsp4 and describes a novel substrate recognition characteristic of EAV nsp4.
CI  - Copyright (c) 2019 American Society for Microbiology.
FAU - Chen, Jiyao
AU  - Chen J
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Wang, Dang
AU  - Wang D
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China wangdang511@126.com 
      vet@mail.hzau.edu.cn.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Sun, Zheng
AU  - Sun Z
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Gao, Li
AU  - Gao L
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Zhu, Xinyu
AU  - Zhu X
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Guo, Jiahui
AU  - Guo J
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Xu, Shangen
AU  - Xu S
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Fang, Liurong
AU  - Fang L
AUID- ORCID: 0000-0003-1406-6409
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Li, Kui
AU  - Li K
AD  - Department of Microbiology, Immunology and Biochemistry, University of Tennessee 
      Health Science Center, Memphis, Tennessee, USA.
FAU - Xiao, Shaobo
AU  - Xiao S
AUID- ORCID: 0000-0003-0023-9188
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China wangdang511@126.com 
      vet@mail.hzau.edu.cn.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Cooperative 
      Innovation Center for Sustainable Pig Production, Wuhan, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190529
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (IKBKG protein, human)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 77238-31-4 (Interferon-beta)
RN  - EC 2.7.11.10 (I-kappa B Kinase)
SB  - IM
MH  - Animals
MH  - Arteriviridae/metabolism
MH  - Arterivirus/metabolism
MH  - Cell Line
MH  - Equartevirus/*metabolism/physiology
MH  - HEK293 Cells
MH  - Horses
MH  - Humans
MH  - I-kappa B Kinase/metabolism/physiology
MH  - Immune Evasion
MH  - Immunity, Innate
MH  - Interferon-beta/*biosynthesis/metabolism
MH  - Porcine respiratory and reproductive syndrome virus/metabolism
MH  - Proteolysis
MH  - Signal Transduction
MH  - Swine
MH  - Viral Nonstructural Proteins/*metabolism
MH  - Virus Replication
PMC - PMC6613749
OTO - NOTNLM
OT  - 3C-like protease
OT  - NF-kappaB essential modulator
OT  - equine arteritis virus
OT  - interferon beta
OT  - porcine reproductive and respiratory syndrome virus
EDAT- 2019/04/05 06:00
MHDA- 2020/06/24 06:00
PMCR- 2019/05/29
CRDT- 2019/04/05 06:00
PHST- 2019/03/04 00:00 [received]
PHST- 2019/03/30 00:00 [accepted]
PHST- 2019/04/05 06:00 [pubmed]
PHST- 2020/06/24 06:00 [medline]
PHST- 2019/04/05 06:00 [entrez]
PHST- 2019/05/29 00:00 [pmc-release]
AID - JVI.00385-19 [pii]
AID - 00385-19 [pii]
AID - 10.1128/JVI.00385-19 [doi]
PST - epublish
SO  - J Virol. 2019 May 29;93(12):e00385-19. doi: 10.1128/JVI.00385-19. Print 2019 Jun 
      15.