Label-free proteomic analysis to confirm the predicted proteome of Corynebacterium pseudotuberculosis under nitrosative stress mediated by nitric oxide

BMC Genomics

Vasco Azevedo⁴  Artur Silva¹  Anderson Miyoshi⁴  Yves Le Loir³  Gustavo HMF Souza²  Edson L Folador⁴  Isabela FS Bastos⁴  Siomar C Soares⁴  Rodrigo D Carvalho⁴  Wanderson M Silva³ 

[1]Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, Pará, Brazil

[2]Waters Corporation, MS Applications and Development Laboratory, São Paulo, Brazil

[3]Agrocampus Ouest, UMR1253 STLO, Rennes 35042, France

[4]Depto de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

关键词: Nitric oxide;    Nitrosative stress;    Label-free proteomics;    Proteomics;    Caseous lymphadenitis;    Corynebacterium pseudotuberculosis;   

Others  :  1089822
DOI  :  10.1186/1471-2164-15-1065

 received in 2014-09-04, accepted in 2014-11-24,  发布年份 2014

【 摘 要 】

Background

Corynebacterium pseudotuberculosis biovar ovis is a facultative intracellular pathogen, and the etiological agent of caseous lymphadenitis in small ruminants. During the infection process, the bacterium is subjected to several stress conditions, including nitrosative stress, which is caused by nitric oxide (NO). In silico analysis of the genome of C. pseudotuberculosis ovis 1002 predicted several genes that could influence the resistance of this pathogen to nitrosative stress. Here, we applied high-throughput proteomics using high definition mass spectrometry to characterize the functional genome of C. pseudotuberculosis ovis 1002 in the presence of NO-donor Diethylenetriamine/nitric oxide adduct (DETA/NO), with the aim of identifying proteins involved in nitrosative stress resistance.

Results

We characterized 835 proteins, representing approximately 41% of the predicted proteome of C. pseudotuberculosis ovis 1002, following exposure to nitrosative stress. In total, 102 proteins were exclusive to the proteome of DETA/NO-induced cells, and a further 58 proteins were differentially regulated between the DETA/NO and control conditions. An interactomic analysis of the differential proteome of C. pseudotuberculosis in response to nitrosative stress was also performed. Our proteomic data set suggested the activation of both a general stress response and a specific nitrosative stress response, as well as changes in proteins involved in cellular metabolism, detoxification, transcriptional regulation, and DNA synthesis and repair.

Conclusions

Our proteomic analysis validated previously-determined in silico data for C. pseudotuberculosis ovis 1002. In addition, proteomic screening performed in the presence of NO enabled the identification of a set of factors that can influence the resistance and survival of C. pseudotuberculosis during exposure to nitrosative stress.

【 授权许可】

   
2014 Silva et al.; licensee BioMed Central.

【 图 表 】

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