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The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line.
Chin Med J (Engl). 2005 Feb 20; 118(4):267-74.CM

Abstract

BACKGROUND

The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) includes sequences encoding the putative protein X4 (ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection.

METHODS

The prokaryotic and eukaryotic protein X4-expressing plasmids were constructed. Recombinant soluble protein X4 was purified from E. coli using ion exchange chromatography, and the preparation was injected into chicken for rising specific polyclonal antibodies. The expression of protein X4 in SARS-CoV-infected Vero E6 cells and lung tissues from patients with SARS was performed using immunofluorescence assay and immunohistochemistry technique. The preliminary function of protein X4 was evaluated by treatment with and over-expression of protein X4 in cell lines. Western blot was employed to evaluate the expression of protein X4 in SARS-CoV particles.

RESULTS

We expressed and purified soluble recombinant protein X4 from E.coli, and generated specific antibodies against protein X4. Western blot proved that the protein X4 was not assembled in the SARS-CoV particles. Indirect immunofluorescence assays revealed that the expression of protein X4 was detected at 8 hours after infection in SARS-CoV-infected Vero E6 cells. It was also detected in the lung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 3T3 cells as determined by cell counting and MTT assays.

CONCLUSION

The results provide the evidence of protein X4 expression following SARS-CoV infection, and may facilitate further investigation of the immunopathological mechanism of SARS.

Authors+Show Affiliations

Center for Human Disease Genomics, Peking University, Beijing 100083, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15740663

Citation

Chen, Ying-yu, et al. "The Protein X4 of Severe Acute Respiratory Syndrome-associated Coronavirus Is Expressed On Both Virus-infected Cells and Lung Tissue of Severe Acute Respiratory Syndrome Patients and Inhibits Growth of Balb/c 3T3 Cell Line." Chinese Medical Journal, vol. 118, no. 4, 2005, pp. 267-74.
Chen YY, Shuang B, Tan YX, et al. The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line. Chin Med J. 2005;118(4):267-74.
Chen, Y. Y., Shuang, B., Tan, Y. X., Meng, M. J., Han, P., Mo, X. N., Song, Q. S., Qiu, X. Y., Luo, X., Gan, Q. N., Zhang, X., Zheng, Y., Liu, S. A., Wang, X. N., Zhong, N. S., & Ma, D. L. (2005). The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line. Chinese Medical Journal, 118(4), 267-74.
Chen YY, et al. The Protein X4 of Severe Acute Respiratory Syndrome-associated Coronavirus Is Expressed On Both Virus-infected Cells and Lung Tissue of Severe Acute Respiratory Syndrome Patients and Inhibits Growth of Balb/c 3T3 Cell Line. Chin Med J. 2005 Feb 20;118(4):267-74. PubMed PMID: 15740663.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line. AU - Chen,Ying-yu, AU - Shuang,Bao, AU - Tan,Ya-xia, AU - Meng,Min-jie, AU - Han,Pu, AU - Mo,Xiao-ning, AU - Song,Quan-sheng, AU - Qiu,Xiao-yan, AU - Luo,Xin, AU - Gan,Qi-ni, AU - Zhang,Xin, AU - Zheng,Ying, AU - Liu,Shun-ai, AU - Wang,Xiao-ning, AU - Zhong,Nan-shan, AU - Ma,Da-long, PY - 2005/3/3/pubmed PY - 2005/4/29/medline PY - 2005/3/3/entrez SP - 267 EP - 74 JF - Chinese medical journal JO - Chin. Med. J. VL - 118 IS - 4 N2 - BACKGROUND: The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) includes sequences encoding the putative protein X4 (ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection. METHODS: The prokaryotic and eukaryotic protein X4-expressing plasmids were constructed. Recombinant soluble protein X4 was purified from E. coli using ion exchange chromatography, and the preparation was injected into chicken for rising specific polyclonal antibodies. The expression of protein X4 in SARS-CoV-infected Vero E6 cells and lung tissues from patients with SARS was performed using immunofluorescence assay and immunohistochemistry technique. The preliminary function of protein X4 was evaluated by treatment with and over-expression of protein X4 in cell lines. Western blot was employed to evaluate the expression of protein X4 in SARS-CoV particles. RESULTS: We expressed and purified soluble recombinant protein X4 from E.coli, and generated specific antibodies against protein X4. Western blot proved that the protein X4 was not assembled in the SARS-CoV particles. Indirect immunofluorescence assays revealed that the expression of protein X4 was detected at 8 hours after infection in SARS-CoV-infected Vero E6 cells. It was also detected in the lung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 3T3 cells as determined by cell counting and MTT assays. CONCLUSION: The results provide the evidence of protein X4 expression following SARS-CoV infection, and may facilitate further investigation of the immunopathological mechanism of SARS. SN - 0366-6999 UR - https://www.unboundmedicine.com/medline/citation/15740663/The_protein_X4_of_severe_acute_respiratory_syndrome_associated_coronavirus_is_expressed_on_both_virus_infected_cells_and_lung_tissue_of_severe_acute_respiratory_syndrome_patients_and_inhibits_growth_of_Balb/c_3T3_cell_line_ L2 - http://www.diseaseinfosearch.org/result/1010 DB - PRIME DP - Unbound Medicine ER -