title: Proteogenomic insights into salt tolerance by a halotolerant alpha-proteobacterium isolated from an Andean saline spring.
abstract: Tistlia consotensis is a halotolerant Rhodospirillaceae that was isolated from a saline spring located in the Colombian Andes with a salt concentration close to seawater (4.5%w/vol). We cultivated this microorganism in three NaCl concentrations, i.e. optimal (0.5%), without (0.0%) and high (4.0%) salt concentration, and analyzed its cellular proteome. For assigning tandem mass spectrometry data, we first sequenced its genome and constructed a six reading frame ORF database from the draft sequence. We annotated only the genes whose products (872) were detected. We compared the quantitative proteome data sets recorded for the three different growth conditions. At low salinity general stress proteins (chaperons, proteases and proteins associated with oxidative stress protection), were detected in higher amounts, probably linked to difficulties for proper protein folding and metabolism. Proteogenomics and comparative genomics pointed at the CrgA transcriptional regulator as a key-factor for the proteome remodeling upon low osmolarity. In hyper-osmotic condition, T. consotensis produced in larger amounts proteins involved in the sensing of changes in salt concentration, as well as a wide panel of transport systems for the transport of organic compatible solutes such as glutamate. We have described here a straightforward procedure in making a new environmental isolate quickly amenable to proteomics.
  
  
  CONCLUSIONS
  The bacterium Tistlia consotensis was isolated from a saline spring in the Colombian Andes and represents an interesting environmental model to be compared with extremophiles or other moderate organisms. To explore the halotolerance molecular mechanisms of the bacterium T. consotensis, we developed an innovative proteogenomic strategy consisting of i) genome sequencing, ii) quick annotation of the genes whose products were detected by mass spectrometry, and iii) comparative proteomics of cells grown in three salt conditions. We highlighted in this manuscript how efficient such an approach can be compared to time-consuming genome annotation when pointing at the key proteins of a given biological question. We documented a large number of proteins found produced in greater amounts when cells are cultivated in either hypo-osmotic or hyper-osmotic conditions. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.
authors.affiliation: Unidad de Saneamiento y Biotecnología Ambiental, Departamento de Biología, Pontificia Universidad Javeriana, POB 56710, Bogotá D.C., Colombia; Colombian Center for Genomics and Bioinformatics of Extreme Environments, GeBiX, Colombia.
authors.forename: Carolina
authors.initials: C
authors.lastname: Rubiano-Labrador
authors.affiliation: CEA, DSV, iBEB, SBTN, Lab Biochim System Perturb, Bagnols-sur-Cèze, F-30207, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Bland
authors.affiliation: CEA, DSV, iBEB, SBTN, Lab Biochim System Perturb, Bagnols-sur-Cèze, F-30207, France.
authors.forename: Guylaine
authors.initials: G
authors.lastname: Miotello
authors.affiliation: CEA, DSV, iBEB, SBTN, Lab Biochim System Perturb, Bagnols-sur-Cèze, F-30207, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Guérin
authors.affiliation: CEA, DSV, iBEB, SBTN, Lab Biochim System Perturb, Bagnols-sur-Cèze, F-30207, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Pible
authors.affiliation: Unidad de Saneamiento y Biotecnología Ambiental, Departamento de Biología, Pontificia Universidad Javeriana, POB 56710, Bogotá D.C., Colombia; Colombian Center for Genomics and Bioinformatics of Extreme Environments, GeBiX, Colombia.
authors.forename: Sandra
authors.initials: S
authors.lastname: Baena
authors.affiliation: CEA, DSV, iBEB, SBTN, Lab Biochim System Perturb, Bagnols-sur-Cèze, F-30207, France. Electronic address: jean.armengaud@cea.fr.
authors.forename: Jean
authors.initials: J
authors.lastname: Armengaud
mesh_terms: D000222:Adaptation, Physiological / Q000502:physiology*; D020561:Alphaproteobacteria / Q000145:classification / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D060985:Natural Springs / Q000382:microbiology*; D016366:Open Reading Frames / Q000502:physiology; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D012965:Sodium Chloride; D014871:Water Microbiology
chemical_list: D001426:Bacterial Proteins; D020543:Proteome; D012965:Sodium Chloride
keywords: Alphaproteobacteria; Draft genome; Halotolerance; Proteogenomics; Rhodospirillaceae