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Identification of cellular proteome using two-dimensional difference gel electrophoresis in ST cells infected with transmissible gastroenteritis coronavirus.

Zhang X, Shi HY, Chen JF, Shi D, Lang HW, Wang ZT, Feng L - Proteome Sci (2013)

Bottom Line: Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs, which is correlated with high morbidity and mortality in suckling piglets.All the protein spots were successfully identified.The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and the mitochondrial pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No, 427 Maduan Street, Nangang District, Harbin 150001, China. fl@hvri.ac.cn.

ABSTRACT

Background: Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs, which is correlated with high morbidity and mortality in suckling piglets. Information remains limited about the comparative protein expression of host cells in response to TGEV infection. In this study, cellular protein response to TGEV infection in swine testes (ST) cells was analyzed, using the proteomic method of two-dimensional difference gel electrophoresis (2D DIGE) coupled with MALDI-TOF-TOF/MS identification.

Results: 33 differentially expressed protein spots, of which 23 were up-regulated and 10 were down-regulated were identified. All the protein spots were successfully identified. The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and the mitochondrial pathway. Western blot analysis was used to validate the changes of alpha tubulin, keratin 19, and prohibitin during TGEV infection.

Conclusions: To our knowledge, we have performed the first analysis of the proteomic changes in host cell during TGEV infection. 17 altered cellular proteins that differentially expressed in TGEV infection were identified. The present study provides protein-related information that should be useful for understanding the host cell response to TGEV infection and the underlying mechanism of TGEV replication and pathogenicity.

No MeSH data available.


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Image of ImageQuant and DeCyder analysis of 35 differentially expressed protein spots in 2D DIGE gels. T + and T- represent TGEV-infected and mock-infected ST cells, respectively.
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Figure 3: Image of ImageQuant and DeCyder analysis of 35 differentially expressed protein spots in 2D DIGE gels. T + and T- represent TGEV-infected and mock-infected ST cells, respectively.

Mentions: To identify cell proteins involved in response to TGEV infection, 2D DIGE proteomics method was performed. Three independent experimental repeat of TGEV-infected and mock-infected ST cells were included in this analysis (Additional file 1: Figure S1). After 2D DIGE, the Cy2, Cy3, and Cy5 channels of each gel were individually imaged, and were analyzed using Decyder software package (version 6.04.11, GE Healthcare). For proteins separated in the pH 4–7 range, 2,710 protein spots were matched. Of which, 33 spots were significantly modified between the TGEV-infected and mock-infected ST cells (2-fold difference in abundance and p < 0.05), including 23 spots that were up-regulated and 10 that were down-regulated (Figure 2 and Figure 3).


Identification of cellular proteome using two-dimensional difference gel electrophoresis in ST cells infected with transmissible gastroenteritis coronavirus.

Zhang X, Shi HY, Chen JF, Shi D, Lang HW, Wang ZT, Feng L - Proteome Sci (2013)

Image of ImageQuant and DeCyder analysis of 35 differentially expressed protein spots in 2D DIGE gels. T + and T- represent TGEV-infected and mock-infected ST cells, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3734006&req=5

Figure 3: Image of ImageQuant and DeCyder analysis of 35 differentially expressed protein spots in 2D DIGE gels. T + and T- represent TGEV-infected and mock-infected ST cells, respectively.
Mentions: To identify cell proteins involved in response to TGEV infection, 2D DIGE proteomics method was performed. Three independent experimental repeat of TGEV-infected and mock-infected ST cells were included in this analysis (Additional file 1: Figure S1). After 2D DIGE, the Cy2, Cy3, and Cy5 channels of each gel were individually imaged, and were analyzed using Decyder software package (version 6.04.11, GE Healthcare). For proteins separated in the pH 4–7 range, 2,710 protein spots were matched. Of which, 33 spots were significantly modified between the TGEV-infected and mock-infected ST cells (2-fold difference in abundance and p < 0.05), including 23 spots that were up-regulated and 10 that were down-regulated (Figure 2 and Figure 3).

Bottom Line: Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs, which is correlated with high morbidity and mortality in suckling piglets.All the protein spots were successfully identified.The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and the mitochondrial pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No, 427 Maduan Street, Nangang District, Harbin 150001, China. fl@hvri.ac.cn.

ABSTRACT

Background: Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs, which is correlated with high morbidity and mortality in suckling piglets. Information remains limited about the comparative protein expression of host cells in response to TGEV infection. In this study, cellular protein response to TGEV infection in swine testes (ST) cells was analyzed, using the proteomic method of two-dimensional difference gel electrophoresis (2D DIGE) coupled with MALDI-TOF-TOF/MS identification.

Results: 33 differentially expressed protein spots, of which 23 were up-regulated and 10 were down-regulated were identified. All the protein spots were successfully identified. The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and the mitochondrial pathway. Western blot analysis was used to validate the changes of alpha tubulin, keratin 19, and prohibitin during TGEV infection.

Conclusions: To our knowledge, we have performed the first analysis of the proteomic changes in host cell during TGEV infection. 17 altered cellular proteins that differentially expressed in TGEV infection were identified. The present study provides protein-related information that should be useful for understanding the host cell response to TGEV infection and the underlying mechanism of TGEV replication and pathogenicity.

No MeSH data available.


Related in: MedlinePlus