1887

Abstract

A proteomic survey of the halophilic archaeon was performed by comparative two-dimensional gel electrophoresis in order to determine the molecular effects of salt stress on the organism. Cells were grown under optimal (2.1 M) and high (3.5 M) NaCl conditions. From this analysis, over 44 protein spots responsive to these conditions were detected. These spots were excised, digested in-gel with trypsin, subjected to QSTAR tandem mass spectrometry (LC/MS/MS) analysis, and identified by comparing the MS/MS-derived peptide sequence to that deduced from the genome. Approximately 40 % of the proteins detected (18 in total) displayed differential abundance based on the detection of at least two peptide fragments per protein and overall MOWSE scores of ≥75 per protein. All of these identified proteins were either uniquely present or 2.3- to 26-fold higher in abundance under one condition compared to the other. The majority of proteins identified in this study were preferentially displayed under optimal salinity and primarily involved in translation, transport and metabolism. However, one protein of interest whose transcript levels were confirmed in these studies to be upregulated under high salt conditions was identified as a homologue of the phage shock protein PspA. The gene belongs to the stress-responsive regulon commonly found among Gram-negative bacteria where its transcription is stimulated by a wide variety of stressors, including heat shock, osmotic shock and prolonged stationary-phase incubation. Homologues of PspA are also found among the genomes of cyanobacteria, higher plants and other , suggesting that this protein may retain some aspects of functional conservation across the three domains of life. Given its integral role in sensing a variety of membrane stressors in bacteria, these results suggest that PspA may play an important role in hypersaline adaptation in .

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2008-05-01
2020-07-10
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References

  1. Baumann P., Qureshi S. A., Jackson S. P.. 1995; Transcription: new insights from studies on Archaea. Trends Genet11:279–283
    [Google Scholar]
  2. Bell S. D., Jackson S.. 1998; Transcription and translation in archaea: a mosaic of eukaryal and bacterial features. Trends Microbiol6:222–228
    [Google Scholar]
  3. Bergler H., Abraham H., Aschauer H., Turnowsky F.. 1994; Inhibition of lipid biosynthesis induces the expression of the pspA gene. Microbiology140:1937–1944
    [Google Scholar]
  4. Besche H., Zwickl P.. 2004; The Thermoplasma acidophilum Lon protease has a Ser-Lys dyad active site. Eur J Biochem271:4361–4365
    [Google Scholar]
  5. Besche H., Tamura N., Tamura T., Zwickl P.. 2004; Mutational analysis of conserved AAA+ residues in the archaeal Lon protease from Thermoplasma acidophilum . FEBS Lett574:161–166
    [Google Scholar]
  6. Bidle K. A.. 2003; Differential expression of genes influenced by changing salinity using RNA arbitrarily primed PCR in the archaeal halophile, Haloferax volcanii . Extremophiles7:1–7
    [Google Scholar]
  7. Bidle K. A., Hanson T. E., Howell K., Nannen J.. 2007; HMG-CoA reductase is regulated by salinity at the level of transcription in Haloferax volcanii . Extremophiles11:49–55
    [Google Scholar]
  8. Brissette J. L., Russel M., Weiner L., Model P.. 1990; Phage shock protein, a stress protein of Escherichia coli . Proc Natl Acad Sci U S A87:862–866
    [Google Scholar]
  9. Brissette J. L., Weiner L., Ripmaster T. L., Model P.. 1991; Characterisation and sequence of the Escherichia coli stress-induced psp operon. J Mol Biol220:35–48
    [Google Scholar]
  10. Daniels C. J., McKee A. H. Z., Doolittle W. F.. 1984; Archaebacterial heat-shock proteins. EMBO J3:745–749
    [Google Scholar]
  11. Darwin A. J.. 2005; The phage-shock-protein response. Mol Microbiol57:621–628
    [Google Scholar]
  12. DeLisa M. P., Lee P., Palmer T., Georgiou G.. 2004; Phage shock protein PspA of Escherichia coli relieves saturation of protein export via the Tat pathway. J Bacteriol186:366–373
    [Google Scholar]
  13. Dennis P. P., Shimmin L. C.. 1997; Evolutionary divergence and salinity-mediated selection in halophilic Archaea. Microbiol Mol Biol Rev61:90–104
    [Google Scholar]
  14. Ferrer C., Mojica F. J. M., Juez G., Rodriguez-Valera F.. 1996; Differentially transcribed regions of Haloferax volcanii genome depending on the medium salinity. J Bacteriol178:309–313
    [Google Scholar]
  15. Gasteiger E., Jung E., Bairoch A.. 2001; SWISS-PROT: connecting biomolecular knowledge via a protein database. Curr Issues Mol Biol3:47–55
    [Google Scholar]
  16. Grant W. D., Gemmell R. T., McGenity T. J.. 1998; Halophiles. In Extremophiles: Microbial Life in Extreme Environments pp93–132 Edited by Horikoshi K., Grant W. D. New York: Wiley and Sons Inc;
    [Google Scholar]
  17. Hankamer B. D., Elderkin S. L., Buck M., Nield J.. 2004; Organization of the AAA(+) adaptor protein PspA is an oligomeric ring. J Biol Chem279:8862–8866
    [Google Scholar]
  18. Izotova L. S., Strongin A. Y., Sterkin V. E., Ostoslavaskaya V. I., Lyublinskaya L. A., Timokhina E. A., Stepanov V. M.. 1983; Purification and properties of serine protease from Halobacterium halobium . J Bacteriol155:826–830
    [Google Scholar]
  19. Javor B.. 1989; Hypersaline Environments: Microbiology and Biogeochemistry Berlin: Springer Verlag;
    [Google Scholar]
  20. Jovanovic G., Weiner L., Model P.. 1996; Identification, nucleotide sequence, and characterization of PspF, the transcriptional activator of the Escherichia coli stress-induced psp operon. J Bacteriol178:1936–1945
    [Google Scholar]
  21. Kirkland P. A., Busby J., Stevens S., Maupin-Furlow J. A.. 2006; Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins. Anal Biochem351:254–259
    [Google Scholar]
  22. Kirkland P. A., Reuter C. J., Maupin-Furlow J. A.. 2007; Effect of proteasome inhibitor clasto -lactacystin- β -lactone on the proteome of the haloarchaeon Haloferax volcanii . Microbiology153:2271–2280
    [Google Scholar]
  23. Kleerebezem M., Tommassen J.. 1993; Expression of the pspA gene stimulates efficient protein export in Escherichia coli . Mol Microbiol7:947–956
    [Google Scholar]
  24. Kleerebezem M., Crielaard W., Tommassen J.. 1996; Involvement of the stress protein PspA (phage shock protein A) of Escherichia coli in maintenance of the proton motive force under stress conditions. EMBO J15:162–171
    [Google Scholar]
  25. Kyrpides N. C., Ouzounis C. A.. 1999; Transcription in Archaea. Proc Natl Acad Sci U S A96:8545–8550
    [Google Scholar]
  26. Langer D., Hain J., Thuriaux P., Zillig W.. 1995; Transcription in Archaea: similarity to that in Eucarya. Proc Natl Acad Sci U S A92:5768–5772
    [Google Scholar]
  27. Livak K. J., Schmittgen T. D.. 2001; Analysis of relative gene expression data using real-time quantitative PCR and the method. Methods25:402–408
    [Google Scholar]
  28. Madern D., Ebel C., Zaccai G.. 2000; Halophilic adaptation of enzymes. Extremophiles4:91–98
    [Google Scholar]
  29. Mojica F. J. M., Cisneros E., Ferrer C., Rodriguez-Valera F., Juez G.. 1997; Osmotically induced response in representatives of halophilic prokaryotes: the bacterium Halomonas elongata and the archeon Haloferax volcanii . J Bacteriol179:5471–5481
    [Google Scholar]
  30. Mullakhanbhai M. F., Larsen H.. 1975; Halobacterium volcanii spec. nov., a Dead Sea halobacterium with a moderate salt requirement. Arch Microbiol104:207–214
    [Google Scholar]
  31. Oren A.. 1998; Microbiology and Biogeochemistry of Hypersaline Environments CRC Press;
    [Google Scholar]
  32. Oren A.. 1999; Bioenergetic aspects of halophilism. Microbiol Mol Biol Rev63:334–348
    [Google Scholar]
  33. Perrière G., Gouy M.. 1996; WWW-Query: an on-line retrieval system for biological sequence banks. Biochimie78:364–369
    [Google Scholar]
  34. Robb F. T., Place A. R., Sowers K. R., Schreier H. J., DasSarma S., Fleischmann E. M.. 1995; Archaea: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  35. Robinson J. L., Pyzyna B., Atrasz R. G.. other authors 2005; Growth kinetics of extremely halophilic Archaea (family Halobacteriaceae) as revealed by Arrhenius plots. J Bacteriol187:923–925
    [Google Scholar]
  36. Rowley G., Spector M., Kormanec J., Roberts M.. 2006; Pushing the envelope: extracytoplasmic stress responses in bacterial pathogens. Nat Rev Microbiol4:383–394
    [Google Scholar]
  37. Soppa J.. 1999; Transcription initiation in Archaea: facts, factors, and future aspects. Mol Microbiol31:1295–1305
    [Google Scholar]
  38. Thomm M.. 1996; Archaeal transcription factors and their role in transcription initiation. FEMS Microbiol Rev18:159–171
    [Google Scholar]
  39. Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G.. 1997; The clustal_x window interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res25:4876–4882
    [Google Scholar]
  40. Weiner L., Model P.. 1994; Role of an Escherichia coli stress-response operon in stationary-phase survival. Proc Natl Acad Sci U S A91:2191–2195
    [Google Scholar]
  41. Weiner L., Brissette J. L., Model P.. 1991; Stress-induced expression of the Escherichia coli phage shock protein operon is dependent on sigma 54 and modulated by positive and negative feedback mechanisms. Genes Dev5:1912–1923
    [Google Scholar]
  42. Wendoloski D., Ferrer C., Dyall-Smith M. L.. 2001; A new simvastatin (mevinolin)-resistance marker from Haloarcula hispanica and a new Haloferax volcanii strain cured of plasmid pHV2. Microbiology147:959–964
    [Google Scholar]
  43. Westphal S., Heins L., Soll J., Vothknecht U. C.. 2001; Vipp1 deletion mutant of Synechocystis : a connection between bacterial phage shock and thylakoid biogenesis?. Proc Natl Acad Sci U S A98:4243–4248
    [Google Scholar]
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