Prohibitin Interacts with envelope proteins of white spot syndrome virus and prevents infection in the red swamp crayfish, Procambarus clarkii.
Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans.
The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education/Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong, China.
Jiang-Feng
JF
Lan
Xin-Cang
XC
Li
Jie-Jie
JJ
Sun
Jing
J
Gong
Xian-Wei
XW
Wang
Xiu-Zhen
XZ
Shi
Li-Jie
LJ
Shi
Yu-Ding
YD
Weng
Xiao-Fan
XF
Zhao
Jin-Xing
JX
Wang
D000818:Animals; D003400:Astacoidea / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D000091402:Prohibitins; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012758:Shellfish / Q000821:virology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D046848:White spot syndrome virus 1 / Q000235:genetics / Q000378:metabolism*
D000091402:Prohibitins; D012097:Repressor Proteins; C548473:VP28 protein, white spot syndrome virus; D014759:Viral Envelope Proteins