PMID- 32038556
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200928
IS  - 1664-302X (Print)
IS  - 1664-302X (Electronic)
IS  - 1664-302X (Linking)
VI  - 10
DP  - 2019
TI  - CD163(DeltaSRCR5) MARC-145 Cells Resist PRRSV-2 Infection via Inhibiting Virus 
      Uncoating, Which Requires the Interaction of CD163 With Calpain 1.
PG  - 3115
LID - 10.3389/fmicb.2019.03115 [doi]
LID - 3115
AB  - Porcine alveolar macrophages without the CD163 SRCR5 domain are resistant to 
      porcine reproductive and respiratory syndrome virus (PRRSV) infection. However, 
      whether the deletion of CD163 SRCR5 in MARC-145 cells confers resistance to PRRSV 
      and interaction of which of the host proteins with CD163 is involved in virus 
      uncoating remain unclear. Here we deleted the SRCR5 domain of CD163 in MARC-145 
      cells using CRISPR/Cas9 to generate a CD163(DeltaSRCR5) MARC-145 cell line. The 
      modification of CD163 had no impact on CD163 expression. CD163(DeltaSRCR5) cells were 
      completely resistant to infection by PRRSV-2 strains Li11, CHR6, TJM, and VR2332. 
      The modified cells showed no cytokine response to PRRSV-2 infection and 
      maintained normal cell vitality comparable with the WT cells. The resistant 
      phenotype of the cells was stably maintained through cell passages. There were no 
      replication transcription complexes in the CD163(DeltaSRCR5) cells. SRCR5 deletion 
      did not disturb the colocalization of CD163 and PRRSV-N in early endosomes (EE). 
      However, the interaction of the viral proteins GP2a, GP3, or GP5 with CD163, 
      which is involved in virus uncoating was affected. Furthermore, 77 CD163-binding 
      cellular proteins affected by the SRCR5 deletion were identified by LC-MS/MS. 
      Inhibition of calpain 1 trapped the virions in EE and forced then into late 
      endosomes but did not block viral attachment and internalization, suggesting that 
      calpain 1 is involved in the uncoating. Overall, CD163(DeltaSRCR5) MARC-145 cells are 
      fully resistant to PRRSV-2 infection and calpain 1 is identified as a novel host 
      protein that interacts with CD163 to facilitate PRRSV uncoating.
CI  - Copyright (c) 2020 Yu, Wei, Dong, Zhu, Zhang, Chen, Liu and Guo.
FAU - Yu, Piao
AU  - Yu P
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Wei, Ruiping
AU  - Wei R
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Dong, Wenjuan
AU  - Dong W
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Zhu, Zhenbang
AU  - Zhu Z
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Zhang, Xiaoxiao
AU  - Zhang X
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Chen, Yaosheng
AU  - Chen Y
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Liu, Xiaohong
AU  - Liu X
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
FAU - Guo, Chunhe
AU  - Guo C
AD  - State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen 
      University, Guangzhou, China.
LA  - eng
PT  - Journal Article
DEP - 20200113
PL  - Switzerland
TA  - Front Microbiol
JT  - Frontiers in microbiology
JID - 101548977
PMC - PMC6990145
OTO - NOTNLM
OT  - CD163
OT  - PRRSV
OT  - SRCR5
OT  - calpain 1
OT  - resistance
EDAT- 2020/02/11 06:00
MHDA- 2020/02/11 06:01
PMCR- 2020/01/13
CRDT- 2020/02/11 06:00
PHST- 2019/11/12 00:00 [received]
PHST- 2019/12/24 00:00 [accepted]
PHST- 2020/02/11 06:00 [entrez]
PHST- 2020/02/11 06:00 [pubmed]
PHST- 2020/02/11 06:01 [medline]
PHST- 2020/01/13 00:00 [pmc-release]
AID - 10.3389/fmicb.2019.03115 [doi]
PST - epublish
SO  - Front Microbiol. 2020 Jan 13;10:3115. doi: 10.3389/fmicb.2019.03115. eCollection 
      2019.