title: Label-free proteomic analysis to confirm the predicted proteome of Corynebacterium pseudotuberculosis under nitrosative stress mediated by nitric oxide.
abstract: 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.
authors.forename: Wanderson M
authors.initials: WM
authors.lastname: Silva
authors.forename: Rodrigo D
authors.initials: RD
authors.lastname: Carvalho
authors.forename: Siomar C
authors.initials: SC
authors.lastname: Soares
authors.forename: Isabela Fs
authors.initials: IF
authors.lastname: Bastos
authors.forename: Edson L
authors.initials: EL
authors.lastname: Folador
authors.forename: Gustavo Hmf
authors.initials: GH
authors.lastname: Souza
authors.forename: Yves
authors.initials: Y
authors.lastname: Le Loir
authors.forename: Anderson
authors.initials: A
authors.lastname: Miyoshi
authors.forename: Artur
authors.initials: A
authors.lastname: Silva
authors.affiliation: Depto de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. vasco@icb.ufmg.br.
authors.forename: Vasco
authors.initials: V
authors.lastname: Azevedo
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D019295:Computational Biology; D003354:Corynebacterium Infections / Q000382:microbiology / Q000662:veterinary; D016925:Corynebacterium pseudotuberculosis / Q000187:drug effects* / Q000302:isolation & purification / Q000378:metabolism; D006041:Goats; D036103:Nanotechnology; D009569:Nitric Oxide / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects; D060066:Protein Interaction Maps; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D020543:Proteome; D009569:Nitric Oxide