Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody

被引：19

作者：

Han, Yuru ^{[1
]}

Zhang, Jiyu ^{[1
]}

Shi, Hongyan ^{[1
]}

Zhou, Ling ^{[2
]}

Chen, Jianfei ^{[1
]}

Zhang, Xin ^{[1
]}

Liu, Jianbo ^{[1
]}

Zhang, Jialin ^{[1
]}

Wang, Xiaobo ^{[1
]}

Ji, Zhaoyang ^{[1
]}

Jing, Zhaoyang ^{[1
]}

Cong, Guangyi ^{[1
]}

Ma, Jingyun ^{[2
]}

Shi, Da ^{[1
]}

Li, Feng ^{[1
]}

机构：

[1] Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Haping Rd 678, Harbin 150069, Heilongjiang, Peoples R China

[2] South China Agr Univ, Coll Anim Sci, Wushan Rd 483, Guangzhou 510642, Guangdong, Peoples R China

来源：

VIRUS RESEARCH | 2019年 / 273卷

基金：

中国国家自然科学基金; 中国博士后科学基金; 黑龙江省自然科学基金;

关键词：

Monoclonal antibody; Swine acute diarrhea syndrome coronavirus; N protein; Epitope mapping; INFECTIOUS-BRONCHITIS VIRUS; SUBCELLULAR-LOCALIZATION; ENTERIC ALPHACORONAVIRUS; TERMINAL DOMAIN; ORGANIZATION; REPLICATION; EXPRESSION; DISCOVERY; SUGGESTS; SEQUENCE;

D O I：

10.1016/j.virusres.2019.197752

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A swine acute diarrhea syndrome coronavirus (SADS-CoV) that causes severe diarrhea in suckling piglets was identified in Southern China in 2017. To develop an antigen that is specific, sensitive, and easy to prepare for serological diagnosis, antigenic sites in the SADS-CoV nucleocapsid (N) protein were screened. We generated and characterized an N-reactive monoclonal antibody (mAb) 3E9 from mice immunized with recombinant N protein. Through fine epitope mapping of mAb 3E9 using a panel of eukaryotic-expressed polypeptides with GFP-tags, we identified the motif (343)DAPVFTPAP(351) as the minimal unit of the linear B-cell epitope recognized by mAb 3E9. Protein sequence alignment indicated that( 343)DAPVFTPAP(351) was highly conserved in different SADS-CoV strains and SADS-related coronaviruses from bat, with one substitution in this motif in HKU2-related bat coronavirus. Using mAb 3E9, we observed that N protein was expressed in the cytoplasm and was in the nucleolus during SADS-CoV replication. N protein was immuno-precipitated from SADS-CoV-infected Vero E6 cells. Taken together, our results indicated that 3E9 mAb could be a useful tool to investigate the structure and function of N protein during viral replication.

引用

页数：7

共 34 条

[1] The nucleoprotein is required for efficient coronavirus genome replication [J].

Almazán, F ;

Galán, C ;

Enjuanes, L .

JOURNAL OF VIROLOGY, 2004, 78 (22) :12683-12688

[2] Modular organization of SARS coronavirus nucleocapsid protein [J].

Chang, CK ;

Sue, SC ;

Yu, TH ;

Hsieh, CM ;

Tsai, CK ;

Chiang, YC ;

Lee, SJ ;

Hsiao, HH ;

Wu, WJ ;

Chang, WL ;

Lin, CH ;

Huang, TH .

JOURNAL OF BIOMEDICAL SCIENCE, 2006, 13 (01) :59-72

[3] The dimer interface of the SARS coronavirus nucleocapsid protein adapts a porcine respiratory and reproductive syndrome virus-like structure [J].

Chang, CK ;

Sue, SC ;

Yu, TH ;

Hsieh, CM ;

Tsai, CK ;

Chiang, YC ;

Lee, SJ ;

Hsiao, HH ;

Wu, WJ ;

Chang, CF ;

Huang, TH .

FEBS LETTERS, 2005, 579 (25) :5663-5668

[4] Antibody detection of SARS-CoV spike and nucleocapsid protein [J].

Chang, MS ;

Lu, YT ;

Ho, ST ;

Wu, CC ;

Wei, TY ;

Chen, CJ ;

Hsu, YT ;

Chu, PC ;

Chen, CH ;

Chu, JM ;

Jan, YL ;

Hung, CC ;

Fan, CC ;

Yang, YC .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 314 (04) :931-936

[5] Structure of the SARS coronavirus nucleocapsid protein RNA-binding dimerization domain suggests a mechanism for helical packaging of viral RNA [J].

Chen, Chun-Yuan ;

Chang, Chung-ke ;

Chang, Yi-Wei ;

Sue, Shih-Che ;

Bai, Hsin-i ;

Riang, Lilianty ;

Hsiao, Chwan-Deng ;

Huang, Tai-huang .

JOURNAL OF MOLECULAR BIOLOGY, 2007, 368 (04) :1075-1086

[6] Biochemical aspects of coronavirus replication and virus-host interaction [J].

Enjuanes, Luis ;

Almazan, Femando ;

Sola, Isabel ;

Zuniga, Sonia .

ANNUAL REVIEW OF MICROBIOLOGY, 2006, 60 :211-230

[7] The nucleocapsid protein of coronavirus infectious bronchitis virus: Crystal structure of its N-terminal domain and multimerization properties [J].

Fan, H ;

Ooi, A ;

Tan, YW ;

Wang, SF ;

Fang, SG ;

Liu, DX ;

Lescar, J .

STRUCTURE, 2005, 13 (12) :1859-1868

[8] Newly emerged porcine enteric alphacoronavirus in southern China: Identification, origin and evolutionary history analysis [J].

Fu, Xinliang ;

Fang, Bo ;

Liu, Yixing ;

Cai, Mengkai ;

Jun, Junming ;

Ma, Jun ;

Bu, Dexin ;

Wang, Lifang ;

Zhou, Pei ;

Wang, Heng ;

Zhang, Guihong .

INFECTION GENETICS AND EVOLUTION, 2018, 62 :179-187

[9]

Galfre G, 1981, Methods Enzymol, V73, P3

[10]

Gong L., 2017, Emerg. Infect. Dis, V23, P1607, DOI DOI 10.3201/EID2309.170915

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