PMID- 32461315
OWN - NLM
STAT- MEDLINE
DCOM- 20201119
LR  - 20210116
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 94
IP  - 15
DP  - 2020 Jul 16
TI  - Structural Characterization of the Helicase nsp10 Encoded by Porcine Reproductive 
      and Respiratory Syndrome Virus.
LID - 10.1128/JVI.02158-19 [doi]
LID - e02158-19
AB  - Currently, an effective therapeutic treatment for porcine reproductive and 
      respiratory syndrome virus (PRRSV) remains elusive. PRRSV helicase nsp10 is an 
      important component of the replication transcription complex that plays a crucial 
      role in viral replication, making nsp10 an important target for drug development. 
      Here, we report the first crystal structure of full-length nsp10 from the 
      arterivirus PRRSV, which has multiple domains: an N-terminal zinc-binding domain 
      (ZBD), a 1B domain, and helicase core domains 1A and 2A. Importantly, our 
      structural analyses indicate that the conformation of the 1B domain from 
      arterivirus nsp10 undergoes a dynamic transition. The polynucleotide substrate 
      channel formed by domains 1A and 1B adopts an open state, which may create enough 
      space to accommodate and bind double-stranded RNA (dsRNA) during unwinding. 
      Moreover, we report a unique C-terminal domain structure that participates in 
      stabilizing the overall helicase structure. Our biochemical experiments also 
      showed that deletion of the 1B domain and C-terminal domain significantly reduced 
      the helicase activity of nsp10, indicating that the four domains must cooperate 
      to contribute to helicase function. In addition, our results indicate that 
      nidoviruses contain a conserved helicase core domain and key amino acid sites 
      affecting helicase function, which share a common mechanism of helicase 
      translocation and unwinding activity. These findings will help to further our 
      understanding of the mechanism of helicase function and provide new targets for 
      the development of antiviral drugs.IMPORTANCE Porcine reproductive and 
      respiratory syndrome virus (PRRSV) is a major respiratory disease agent in pigs 
      that causes enormous economic losses to the global swine industry. PRRSV helicase 
      nsp10 is a multifunctional protein with translocation and unwinding activities 
      and plays a vital role in viral RNA synthesis. Here, we report the first 
      structure of full-length nsp10 from the arterivirus PRRSV at 3.0-A resolution. 
      Our results show that the 1B domain of PRRSV nsp10 adopts a novel open state and 
      has a unique C-terminal domain structure, which plays a crucial role in nsp10 
      helicase activity. Furthermore, mutagenesis and structural analysis revealed 
      conservation of the helicase catalytic domain across the order Nidovirales 
      (families Arteriviridae and Coronaviridae). Importantly, our results will provide 
      a structural basis for further understanding the function of helicases in the 
      order Nidovirales.
CI  - Copyright (c) 2020 American Society for Microbiology.
FAU - Shi, Yuejun
AU  - Shi Y
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Tong, Xiaohan
AU  - Tong 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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Ye, Gang
AU  - Ye G
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Xiu, Ruixue
AU  - Xiu R
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Li, Lisha
AU  - Li 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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Sun, Limeng
AU  - Sun 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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Shi, Jiale
AU  - Shi 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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Li, Mengxia
AU  - Li M
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Song, Yunfeng
AU  - Song Y
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Fan, Chengpeng
AU  - Fan C
AD  - Department of Biochemistry and Molecular Biology, School of Basic Medical 
      Sciences, Wuhan University, Wuhan, China.
FAU - Shi, Ke
AU  - Shi K
AD  - Department of Biochemistry, Molecular Biology, and Biophysics, University of 
      Minnesota, Minneapolis, Minnesota, USA.
FAU - Fu, Zhen F
AU  - Fu ZF
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, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
AD  - Department of Pathology, College of Veterinary Medicine, University of Georgia, 
      Athens, Georgia, 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.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FAU - Peng, Guiqing
AU  - Peng G
AD  - State Key Laboratory of Agricultural Microbiology, College of Veterinary 
      Medicine, Huazhong Agricultural University, Wuhan, China penggq@mail.hzau.edu.cn.
AD  - Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The 
      Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200716
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (RNA, Double-Stranded)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Proteins)
RN  - EC 3.6.4.13 (RNA Helicases)
SB  - IM
MH  - Porcine respiratory and reproductive syndrome virus/*enzymology/genetics
MH  - Protein Domains
MH  - RNA Helicases/*chemistry/genetics
MH  - RNA, Double-Stranded/*chemistry/genetics
MH  - RNA, Viral/*chemistry/genetics
MH  - Viral Proteins/*chemistry/genetics
PMC - PMC7375384
OTO - NOTNLM
OT  - helicase
OT  - nonstructural protein 10
OT  - porcine reproductive and respiratory syndrome virus
OT  - structure
EDAT- 2020/05/29 06:00
MHDA- 2020/11/20 06:00
PMCR- 2021/01/16
CRDT- 2020/05/29 06:00
PHST- 2019/12/25 00:00 [received]
PHST- 2020/05/20 00:00 [accepted]
PHST- 2020/05/29 06:00 [pubmed]
PHST- 2020/11/20 06:00 [medline]
PHST- 2020/05/29 06:00 [entrez]
PHST- 2021/01/16 00:00 [pmc-release]
AID - JVI.02158-19 [pii]
AID - 02158-19 [pii]
AID - 10.1128/JVI.02158-19 [doi]
PST - epublish
SO  - J Virol. 2020 Jul 16;94(15):e02158-19. doi: 10.1128/JVI.02158-19. Print 2020 Jul 
      16.