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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.    Electronic supplementary material  The online version of this article (doi:10.1186/1471-2164-15-1065) contains supplementary material, which is available to authorized users.

bmc genomics

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