猪传染性胃肠炎病毒Nsp2与宿主细胞蛋白PSMD11相互作用的研究

doi:10.16431/j.cnki.1671-7236.2021.07.033

Abstract

Abstract: To determine whether there was an interaction between Transmissible gastroenteritis virus (TGEV) nonstructural protein 2 (Nsp2) and the host cell protein 26S proteasome non-ATPase regulatory subunit 11 (PSMD11),RT-PCR was used to amplify the porcine PSMD11 gene and eukaryotic expression vector pCMV-Myc-PSMD11 was constructed,which was verified by sequencing and double enzyme digestion.And then pCMV-Myc-PSMD11 was transfected into intestinal epithelial cell lines J2 (IPEC-J2),Western blotting and indirect immunofluorescence assay (IFA) were used to detect whether pCMV-Myc-PSMD11 could be expressed in IPEC-J2 cells.Then,the co-immunoprecipitation (Co-IP) experiment was used to determine the interaction between TGEV Nsp2 and PSMD11 of the host cell,and the co-localization of TGEV Nsp2 and PSMD11 in the host cell was observed by laser confocal microscope.The results showed that porcine PSMD11 gene was successfully amplified,with a size of 1 474 bp,and the gene sequence was completely consistent with the standard sequence.The constructed eukaryotic expression vector pCMV-Myc-PSMD11 could successfully express PSMD11 protein in IPEC-J2 cells.The results of Co-IP indicated that there was an interaction between PSMD11 and Nsp2.The results of the colocalization experiment showed that the interaction between PSMD11 and Nsp2 occured in the cytoplasm,and the expression position of PSMD11 in the cell was not changed due to the expression of Nsp2.The results of this study provided new clues for further study of the important role of TGEV Nsp2 in the process of virus infection.

Key words: Transmissible gastroenteritis virus (TGEV); Nsp2; PSMD11; protein interaction

CLC Number:

S852.65⁺1

SUN Aoying, WANG Yanan, JIANG Yanping, CUI Wen, QIAO Xinyuan, ZHOU Han, TANG Lijie, XU Yigang, LI Yijing, WANG Li. Study on Interaction Between Transmissible Gastroenteritis Virus Nsp2 and Host Cell Protein PSMD11[J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(7): 2569-2576.

References

[1] 焦韵洁,张欣,朱海鹏,等.猪传染性胃肠炎病毒可视化RT-LAMP检测方法的建立[J].中国兽医学报,2020,40(7):1272-1279. JIAO Y J,ZHANG X,ZHU H P,et al.Establishment of a visual RT-LAMP detection assay for TGEV[J].Chinese Journal of Veterinary Science,2020,40(7):1272-1279.(in Chinese)
[2] ZHOU J,HUANG F,HUA X G,et al.Inhibition of porcine Transmissible gastroenteritis virus (TGEV) replication in Mini-pigs by shRNA[J].Virus Research,2010,149(1):51-55.
[3] CHATTHA K,ROTH J,SAIF L.Strategies for design and application of enteric viral vaccines[J].Annual Review of Animal Biosciences,2014,3(1):375-395.
[4] KWONIL J,HUI H,BRYAN E,et al.Pathogenicity of 2 Porcine deltacoronavirus strains in gnotobiotic pigs[J].Emerging Infectious Diseases,2015,21(4):650-654.
[5] JUNG K,WANG Q,SCHEUER K,et al.Pathology of US Porcine epidemic diarrhea virus strain PC21A in gnotobiotic pigs[J].Emerging Infectious Diseases,2014,20(4):668-671.
[6] WANG L,QIAO X,ZHANG S,et al.Porcine transmissible gastroenteritis virus nonstructural protein 2 contributes to inflammation via NF-κB activation[J].Virulence,2018,9(1):1685-1698.
[7] GUMA M,STEPNIAK D,SHAKED H,et al.Constitutive intestinal NF-κB does not trigger destructive inflammation unless accompanied by MAPK activation[J].Journal of Experimental Medicine,2011,208(9):1889-1900.
[8] GHOSH S K.Shared principles in NF-kappa B signaling[J].Cell,2008,132(3):344-362.
[9] HAYDEN M S,GHOSH S.NF-κB in immunobiology[J].Cell Research,2011,21(2):223-244.
[10] 王亚楠,韩露露,孙傲颖,等.宿主细胞蛋白HSPA2与TGEV Nsp2的相互作用及其对病毒复制的影响[J].黑龙江畜牧兽医,2020,21:1-6. WANG Y N,HAN L L,SUN A Y,et al.Interaction between host cell protein HSPA2 and TGEV Nsp2 and its effect on virus replication[J].Heilongjiang Animal Science and Veterinary Medicine,2020,21:1-6.(in Chinese)
[11] VOGES D,ZWICKL P,BAUMEISTER W.The 26S proteasome:A molecular machine designed for controlled proteolysis[J].Annual Review of Biochemistry,1999,68(1):1015-1068.
[12] 段群棚,李晓玉,赵硕,等.2016-2019年广西部分猪场仔猪病毒性腹泻的流行病学调查[J].中国畜牧兽医,2020,47(2):564-574. DUAN Q P,LI X Y,ZHAO S,et al.Epidemiological investigation on diarrhea viral disease of piglets in some pig farms in Guangxi during 2016 to 2019[J].China Animal Husbandry & Veterinary Medicine,2020,47(2):564-574.(in Chinese)
[13] 王睿敏.猪四种腹泻病毒快速鉴别检测方法及猪流行性腹泻病毒分子流行病学研究[D].南宁:广西大学,2019. WANG R M.Study on the rapid differential methods for detection of four porcine diarrhea viruses and molecular epidemiology of porcine epidemic diarrhea virus[D].Nanning:Guangxi University,2019.(in Chinese)
[14] 陶嘉磊.基于Ⅰ型干扰素信号通路探讨清肺口服液类黄酮组分抗RSV研究[D].南京:南京中医药大学,2020. TAO J L.Study on anti-RSV of flavonoid components in Qingfei oral liquid based on the type Ⅰ interferon signaling pathway[D].Nanjing:Nanjing University of Chinese Medicine,2020.(in Chinese)
[15] WANG R,YU R,CHEN B,et al.Identification of host cell proteins that interact with the M protein of Porcine epidemic diarrhea virus[J].Veterinary Microbiology,2020,246:108729.
[16] WANG L,ZHOU L,ZHANG H,et al.Interactome profile of the host cellular proteins and the nonstructural protein 2 of Porcine reproductive and respiratory syndrome virus[J].PLoS One,2014,9(6):e99176.
[17] 李朝华.免疫共沉淀联合质谱筛选与HCMV IE86相互作用的蛋白[D].青岛:青岛大学,2018. LI C H.Identification of HCMV IE86 protein complex by coimmunoprecipitation and LC-MS/MS analysis[D].Qingdao:Qingdao University,2018.(in Chinese)
[18] USHIJIMA Y,KOSHIZUKA T,GOSHIMA F,et al.Herpes simplex virus type 2 UL56 interacts with the ubiquitin ligase Nedd4 and increases its ubiquitination[J].Journal of Virology,2008,82(11):5220-5233.
[19] 王书文,吕闯,蔡雪辉.伪狂犬病病毒UL56蛋白与宿主蛋白Nedd4相互作用的研究[J].中国预防兽医学报,2019,41(6):553-559. WANG S W,LYU C,CAI X H.PRV protein UL56 interacts with Nedd4[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(6):553-559.(in Chinese)
[20] 王乐乐.应用高三尖杉酯碱抑制蛋白合成可在胰腺癌细胞中通过激活MEK1/ERK1/2信号途径诱导PSMD11蛋白的快速合成[D].济南:山东大学,2018. WANG L L.Homoharringtonine could induce quick protein synthesis of PSMD11 through activating MEK1/ERK1/2 signaling pathway in pancreatic cancer cells[D].Jinan:Shandong University,2018.(in Chinese)
[21] 孙建.去泛素化酶USP8对缝隙连接蛋白CX43的去泛素化作用和对其功能的影响[D].上海:上海交通大学,2018. SUN J.The deubquitination of USP8 on CX43 and the influence on function of CX43[D].Shanghai:Shanghai Jiao Tong University,2018.(in Chinese)
[22] 王洋,金一.泛素蛋白酶体途径及其在体外受精中的作用研究进展[J].中国畜牧兽医,2018,45(2):338-348. WANG Y,JIN Y.Research progress on ubiquitin proteasome pathway and its function in vitro fertilization[J].China Animal Husbandry & Veterinary Medicine,2018,45(2):338-348.(in Chinese)
[23] 程爽.泛素蛋白酶体途径在猪圆环病毒2型体外感染中的作用研究[D].武汉:华中农业大学,2014. CHENG S.Role of ubiquitin-proteasome system in the Porcine circovirus type 2 infection in vitro[D].Wuhan:Huazhong Agricultural University,2014.(in Chinese)
[24] CHOI A G,WONG J,MARCHANT D,et al.The ubiquitin-proteasome system in positive-strand RNA virus infection[J].Reviews in Medical Virology,2013,23(2):85-96.
[25] WEI H,LI J Z,LI Y H,et al.microRNA-451 inhibits inflammation and proliferation of glomerular mesangial cells through down-regulating PSMD11 and NF-κB p65[J].Bioscience Reports,2019,39(10):BSR20191455.
[26] WEI B,LI H Z,ZHI F Z,et al.Investigations into the role of 26S proteasome non-ATPase regulatory subunit 13 in neuroinflammation[J].Neuroimmunomo-dulation,2014,21(6):331-337.

Study on Interaction Between Transmissible Gastroenteritis Virus Nsp2 and Host Cell Protein PSMD11

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