1887

Abstract

CgtA is a member of the Obg/Gtp1 subfamily of small GTP-binding proteins. CgtA homologues have been found in various prokaryotic and eukaryotic organisms, ranging from bacteria to humans. Nevertheless, despite the fact that is an essential gene in most bacterial species, its function in the regulation of cellular processes is largely unknown. Here it has been demonstrated that in two bacterial species, and , the gene product enhances survival of cells after UV irradiation. Expression of the gene was found to be enhanced after UV irradiation of both and . Moderate overexpression of resulted in higher UV resistance of wild-type and strains, but not in , , and mutant hosts. Overexpression of the gene in the mutant, which is very sensitive to UV light, restored the level of survival of UV-irradiated cells to the levels observed for wild-type bacteria. Moreover, the basal level of the RecA protein was lower in a temperature-sensitive mutant of than in the strain, and contrary to wild-type bacteria, no significant increase in gene expression was observed after UV irradiation of this mutant. Finally, stimulation of gene transcription under these conditions was impaired in the mutant. All these results strongly suggest that the gene product is involved in DNA repair processes, most probably by stimulation of gene expression and resultant activation of RecA-dependent DNA repair pathways.

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2003-07-01
2020-03-31
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References

  1. Arigoni F., Talabot F., Peitsch M., Edgerton M. D., Meldrum E., Allet E., Fish R., Jamotte T., Curchod M.-L., Loferer H.. 1998; A genome-based approach for the identification of essential bacterial genes. Nat Biotechnol16:851–856
    [Google Scholar]
  2. Belas R., Mileham A., Cohn D., Hilmen M., Simon M., Silverman M.. 1982; Bacterial luminescence: isolation and expression of the luciferase genes from Vibrio harveyi . Science218:791–793
    [Google Scholar]
  3. Buglino J., Shen V., Hakimian P., Lima C. D.. 2002; Structural and biochemical analysis of the Obg GTP binding protein. Structure10:1581–1592
    [Google Scholar]
  4. Chang A. C. Y., Cohen S. N.. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol134:1141–1156
    [Google Scholar]
  5. Courcelle J., Khodursky A., Peter B., Brown P. O., Hanawalt P. C.. 2001; Comparative gene expression profiles following UV exposure in wild-type and SOS-deficient Escherichia coli . Genetics158:41–64
    [Google Scholar]
  6. Czyż A., Wróbel B., Wȩgrzyn G.. 2000a; Vibrio harveyi bioluminescence plays a role in stimulation of DNA repair. Microbiology146:283–288
    [Google Scholar]
  7. Czyż A., Jasiecki J., Bogdan A., Szpilewska H., Wȩgrzyn G.. 2000b; Genetically modified Vibrio harveyi strains as potential bioindicators of mutagenic pollution of marine environments. Appl Environ Microbiol66:599–605
    [Google Scholar]
  8. Czyż A., Zielke R., Konopa G., Wȩgrzyn G.. 2001; A Vibrio harveyi insertional mutant in the cgtA ( obg , yhbZ ) gene, whose homologues are present in diverse organisms ranging from bacteria to humans and are essential genes in many bacterial species. Microbiology147:183–191
    [Google Scholar]
  9. Czyż A., Szpilewska H., Dutkiewicz R., Kowalska W., Biniewska-Godlewska A., Wȩgrzyn G.. 2002; Comparison of the Ames test and a newly developed assay for detection of mutagenic pollution of marine environments. Mutat Res519:67–74
    [Google Scholar]
  10. Durland R. H., Toukdarian A., Fang F., Helinski D. R.. 1990; Mutations in the trfA replication gene of the broad-host range plasmid RK2 results in elevated plasmid copy number. J Bacteriol172:3859–3867
    [Google Scholar]
  11. Dutkiewicz R., Słomińska M., Wȩgrzyn G., Czyż A.. 2002; Overexpression of the cgtA ( yhbZ , obgE ) gene, coding for an essential GTP-binding protein, impairs the regulation of chromosomal functions in Escherichia coli . Curr Microbiol45:440–445
    [Google Scholar]
  12. Jensen K. F.. 1993; The Escherichia coli ‘wild types' W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol175:3401–3407
    [Google Scholar]
  13. Klein G., Z˙mijewski M., Krzewska J., Czeczatka M., Lipińska B.. 1998; Cloning and characterization of the dnaK heat shock operon of the marine bacterium Vibrio harveyi . Mol Gen Genet259:179–189
    [Google Scholar]
  14. Kobayashi G., Moriya S., Wada C.. 2001; Deficiency of essential GTP-binding protein ObgE in Escherichia coli inhibits chromosome partition. Mol Microbiol41:1037–1051
    [Google Scholar]
  15. Kok J., Trach K. A., Hoch J. A.. 1994; Effects on Bacillus subtilis of a conditional lethal mutation in the essential GTP-binding protein Obg. J Bacteriol176:7155–7160
    [Google Scholar]
  16. Lin B., Maddock J. R.. 2001; The N-terminal domain of the Caulobacter crescentus CgtA protein does not function as a guanine nucleotide exchange factor. FEBS Lett489:108–111
    [Google Scholar]
  17. Lin B., Covalle K. L., Maddock J. R.. 1999; The Caulobacter crescentus CgtA protein displays unusual guanine nucleotide binding and exchange properties. J Bacteriol181:5825–5832
    [Google Scholar]
  18. Lin B., Skidmore J. M., Bhatt A., Pfeffer S. M., Pawloski L., Maddock J. R.. 2001; Alanine scan mutagenesis of the switch I domain the Caulobacter crescentus CgtA protein reveals critical amino acids required for in vivo function. Mol Microbiol39:924–934
    [Google Scholar]
  19. Linn S.. 1996; The DNases, topoisomerases and helicases of Escherichia coli . In Escherichia coli and Salmonella: Cellular and Molecular Biology . pp 764–772 Edited by Neidhardt F. C., Curtiss R. III, Ingraham J. L., Lin E. C. C., Low K. B., Magasanik B., Reznikoff W. S., Riley M., Schaechter M., Umbarger H. E.. Washington, DC: American Society for Microbiology;
  20. MacKenzie C., Chidambaram M., Sodergren E. J., Kaplan S., Weinstock G. M.. 1995; DNA repair mutants of Rhodobacter sphaeroides . J Bacteriol177:3027–3035
    [Google Scholar]
  21. Maddock J., Bhatt A., Koch M., Skidmore J.. 1997; Identification of an essential Caulobacter crescentus gene encoding a member of the Obg family of GTP-binding proteins. J Bacteriol179:6426–6431
    [Google Scholar]
  22. Miyamoto C. M., Lin Y. H., Meighen E. A.. 2000; Control of bioluminescence in Vibrio fischeri by the LuxO signal response regulator. Mol Microbiol36:594–607
    [Google Scholar]
  23. Rupp W. D., Curtiss R. III. 1996; DNA repair mechanisms. In Escherichia coli and Salmonella: Cellular and Molecular Biology . pp 2277–2294 Edited by Neidhardt F. C., Ingraham J. L., Lin E. C. C., Low K. B., Magasanik B., Reznikoff W. S., Riley M., Schaechter M., Umbarger H. E.. Washington, DC: American Society for Microbiology;
  24. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  25. Scott J. M., Haldenwang W. G.. 1999; Obg, an essential GTP binding protein of Bacillus subtilis , is necessary for stress activation of transcription factor σ B. J Bacteriol181:4653–4660
    [Google Scholar]
  26. Scott J. M., Ju J., Mitchell T., Haldenwang W. G.. 2000; The Bacillus subtilis GTP-binding protein Obg and regulators of the σ B stress response transcription factor cofractionate with ribosomes. J Bacteriol182:2771–2777
    [Google Scholar]
  27. Sikora-Borgula A., Słomińska M., Trzonkowski P., Zielke R., Myśliwski A., Wȩgrzyn G., Czyż A.. 2002; A role for the common GTP-binding protein in coupling of chromosome replication to cell growth and cell division. Biochem Biophys Res Commun292:333–338
    [Google Scholar]
  28. Singer M., Baker T. A., Schnitzler G.. 7 other authors 1989; A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli . Microbiol Rev53:1–24
    [Google Scholar]
  29. Słomińska M., Konopa G., Wȩgrzyn G., Czyż A.. 2002; Impaired chromosome partitioning and synchronization of DNA replication initiation in a Vibrio harveyi insertional mutant in the cgtA gene coding for a common GTP-binding protein. Biochem J362:579–584
    [Google Scholar]
  30. Taft-Benz S. A., Schaaper R. M.. 1999; The C-terminal domain of DnaQ contains the polymerase binding site. J Bacteriol181:2963–2965
    [Google Scholar]
  31. Tan J., Jakob U., Bardwell J. C.. 2002; Overexpression of two different GTPases rescues a null mutation in a heat-induced rRNA methyltransferase. J Bacteriol184:2692–2698
    [Google Scholar]
  32. Vidwans S. J., Ireton K., Grossman A. D.. 1995; Possible role for the essential GTP-binding protein Obg in regulating the initiation of sporulation in Bacillus subtilis . J Bacteriol177:3308–3311
    [Google Scholar]
  33. Vieira J., Messing J.. 1982; The pUC plasmids, an M13mp7-derived system for insertional mutagenesis and sequencing with synthetic universal primers. Gene19:259–268
    [Google Scholar]
  34. Walker G. C.. 1996; The SOS response of Escherichia coli . In Escherichia coli and Salmonella: Cellular and Molecular Biology , pp. 1400–1416 Edited by Neidhardt F. C., Curtiss R. III, Ingraham J. L., Lin E. C. C., Low K. B., Magasanik B., Reznikoff W. S., Riley M., Schaechter M., Umbarger H. E. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  35. Wȩgrzyn G., Taylor K.. 1992; Inheritance of the replication complex by one of two daughter copies during λ plasmid replication in Escherichia coli . J Mol Biol226:681–688
    [Google Scholar]
  36. Wȩgrzyn A., Wȩgrzyn G., Taylor K.. 1995a; Protection of coliphage λ O initiator protein from proteolysis in the assembly of the replication complex in vivo . Virology207:179–184
    [Google Scholar]
  37. Wȩgrzyn G., Wȩgrzyn A., Konieczny I., Bielawski K., Konopa G., Obuchowski M., Helinski D. R., Taylor K.. 1995b; Involvement of the host initiator function dnaA in the replication of coliphage λ . Genetics139:1469–1481
    [Google Scholar]
  38. Wittinghofer A.. 2002; Obg, a G domain with a beautiful extension. Structure10:1471–1472
    [Google Scholar]
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