Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Jun;177(11):3027–3035. doi: 10.1128/jb.177.11.3027-3035.1995

DNA repair mutants of Rhodobacter sphaeroides.

C Mackenzie 1, M Chidambaram 1, E J Sodergren 1, S Kaplan 1, G M Weinstock 1
PMCID: PMC176989  PMID: 7768798

Abstract

The genome of the photosynthetic eubacterium Rhodobacter sphaeroides 2.4.1 comprises two chromosomes and five endogenous plasmids and has a 65% G+C base composition. Because of these characteristics of genome architecture, as well as the physiological advantages that allow this organism to live in sunlight when in an anaerobic environment, the sensitivity of R. sphaeroides to UV radiation was compared with that of the more extensively studied bacterium Escherichia coli. R. sphaeroides was found to be more resistant, being killed at about 60% of the rate of E. coli. To begin to analyze the basis for this increased resistance, a derivative of R. sphaeroides, strain 2.4.1 delta S, which lacks the 42-kb plasmid, was mutagenized with a derivative of Tn5, and the transposon insertion mutants were screened for increased UV sensitivity (UVs). Eight UVs strains were isolated, and the insertion sites were determined by contour-clamped homogeneous electric field pulsed-field gel electrophoresis. These mapped to at least five different locations in chromosome I. Preliminary analysis suggested that these mutants were deficient in the repair of DNA damage. This was confirmed for three loci by DNA sequence analysis, which showed the insertions to be within genes homologous to uvrA, uvrB, and uvrC, the subunits of the nuclease responsible for excising UV damage.

Full Text

The Full Text of this article is available as a PDF (404.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abeliovich A., Kaplan S. Bacteriophages of Rhodopseudomonas spheroides: isolation and characterization of a Rhodopseudomonas spheroides bacteriophage. J Virol. 1974 Jun;13(6):1392–1399. doi: 10.1128/jvi.13.6.1392-1399.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abratt V. R., Jones D. T., Woods D. R. Isolation and physiological characterization of mitomycin C-sensitive/UV-sensitive mutants in Bacteroides fragilis. J Gen Microbiol. 1985 Sep;131(9):2479–2483. doi: 10.1099/00221287-131-9-2479. [DOI] [PubMed] [Google Scholar]
  3. Antopol S. C., Ellner P. D. Susceptibility of Legionella pneumophila to ultraviolet radiation. Appl Environ Microbiol. 1979 Aug;38(2):347–348. doi: 10.1128/aem.38.2.347-348.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arrage A. A., Phelps T. J., Benoit R. E., White D. C. Survival of subsurface microorganisms exposed to UV radiation and hydrogen peroxide. Appl Environ Microbiol. 1993 Nov;59(11):3545–3550. doi: 10.1128/aem.59.11.3545-3550.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barbé J., Gibert I., Llagostera M., Guerrero R. DNA repair systems in the phototrophic bacterium Rhodobacter capsulatus. J Gen Microbiol. 1987 Apr;133(4):961–966. doi: 10.1099/00221287-133-4-961. [DOI] [PubMed] [Google Scholar]
  6. Brash D. E., Haseltine W. A. UV-induced mutation hotspots occur at DNA damage hotspots. Nature. 1982 Jul 8;298(5870):189–192. doi: 10.1038/298189a0. [DOI] [PubMed] [Google Scholar]
  7. Butler R. C., Lund V., Carlson D. A. Susceptibility of Campylobacter jejuni and Yersinia enterocolitica to UV radiation. Appl Environ Microbiol. 1987 Feb;53(2):375–378. doi: 10.1128/aem.53.2.375-378.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Calero S., Fernandez de Henestrosa A. R., Barbé J. Molecular cloning, sequence and regulation of expression of the recA gene of the phototrophic bacterium Rhodobacter sphaeroides. Mol Gen Genet. 1994 Jan;242(1):116–120. doi: 10.1007/BF00277356. [DOI] [PubMed] [Google Scholar]
  9. Campbell L. A., Yasbin R. E. Deoxyribonucleic acid repair capacities of Neisseria gonorrhoeae: absence of photoreactivation. J Bacteriol. 1979 Dec;140(3):1109–1111. doi: 10.1128/jb.140.3.1109-1111.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Choudhary M., Mackenzie C., Nereng K. S., Sodergren E., Weinstock G. M., Kaplan S. Multiple chromosomes in bacteria: structure and function of chromosome II of Rhodobacter sphaeroides 2.4.1T. J Bacteriol. 1994 Dec;176(24):7694–7702. doi: 10.1128/jb.176.24.7694-7702.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chung C. T., Niemela S. L., Miller R. H. One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2172–2175. doi: 10.1073/pnas.86.7.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. David H. L. Response of Mycobacteria to ultraviolet light radiation. Am Rev Respir Dis. 1973 Nov;108(5):1175–1185. doi: 10.1164/arrd.1973.108.5.1175. [DOI] [PubMed] [Google Scholar]
  13. Donohue T. J., Chory J., Goldsand T. E., Lynn S. P., Kaplan S. Structure and physical map of Rhodopseudomonas sphaeroides bacteriophage RS1 DNA. J Virol. 1985 Jul;55(1):147–157. doi: 10.1128/jvi.55.1.147-157.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Donohue T. J., Kaplan S. Genetic techniques in rhodospirillaceae. Methods Enzymol. 1991;204:459–485. doi: 10.1016/0076-6879(91)04024-i. [DOI] [PubMed] [Google Scholar]
  15. Fernandez de Henestrosa A. R., Barbé J. Induction of the alkA gene of Escherichia coli in gram-negative bacteria. J Bacteriol. 1991 Dec;173(23):7736–7740. doi: 10.1128/jb.173.23.7736-7740.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fernandez de Henestrosa A. R., Calero S., Barbé J. Expression of the recA gene of Escherichia coli in several species of gram-negative bacteria. Mol Gen Genet. 1991 May;226(3):503–506. doi: 10.1007/BF00260664. [DOI] [PubMed] [Google Scholar]
  17. Friedman B. M., Yasbin R. E. The genetics and specificity of the constitutive excision repair system of Bacillus subtilis. Mol Gen Genet. 1983;190(3):481–486. doi: 10.1007/BF00331080. [DOI] [PubMed] [Google Scholar]
  18. Gasc A. M., Sicard N., Claverys J. P., Sicard A. M. Lack of SOS repair in Streptococcus pneumoniae. Mutat Res. 1980 Apr;70(2):157–165. doi: 10.1016/0027-5107(80)90155-4. [DOI] [PubMed] [Google Scholar]
  19. Gomez-Eichelmann M. C., Levy-Mustri A., Ramirez-Santos J. Presence of 5-methylcytosine in CC(A/T)GG sequences (Dcm methylation) in DNAs from different bacteria. J Bacteriol. 1991 Dec;173(23):7692–7694. doi: 10.1128/jb.173.23.7692-7694.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gopalakrishnan A. S., Chen Y. C., Temkin M., Dowhan W. Structure and expression of the gene locus encoding the phosphatidylglycerophosphate synthase of Escherichia coli. J Biol Chem. 1986 Jan 25;261(3):1329–1338. [PubMed] [Google Scholar]
  21. Harm W. Biological determination of the germicidal activity of sunlight. Radiat Res. 1969 Oct;40(1):63–69. [PubMed] [Google Scholar]
  22. Hempel J., Nicholas H., Lindahl R. Aldehyde dehydrogenases: widespread structural and functional diversity within a shared framework. Protein Sci. 1993 Nov;2(11):1890–1900. doi: 10.1002/pro.5560021111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hennecke F., Kolmar H., Bründl K., Fritz H. J. The vsr gene product of E. coli K-12 is a strand- and sequence-specific DNA mismatch endonuclease. Nature. 1991 Oct 24;353(6346):776–778. doi: 10.1038/353776a0. [DOI] [PubMed] [Google Scholar]
  24. Howard-Flanders P. DNA repair. Annu Rev Biochem. 1968;37:175–200. doi: 10.1146/annurev.bi.37.070168.001135. [DOI] [PubMed] [Google Scholar]
  25. Lieb M. Spontaneous mutation at a 5-methylcytosine hotspot is prevented by very short patch (VSP) mismatch repair. Genetics. 1991 May;128(1):23–27. doi: 10.1093/genetics/128.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lo R. Y., MacDonald L. E. Pasteurella haemolytica is highly sensitive to ultraviolet irradiation. Mutat Res. 1991 Jul;263(3):159–163. doi: 10.1016/0165-7992(91)90056-a. [DOI] [PubMed] [Google Scholar]
  27. Majumdar S., Chandra A. K. UV-repair and mutagenesis in Azotobacter vinelandii. I. Repair of UV-induced damages. Zentralbl Mikrobiol. 1985;140(3):247–254. [PubMed] [Google Scholar]
  28. Minton K. W. DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans. Mol Microbiol. 1994 Jul;13(1):9–15. doi: 10.1111/j.1365-2958.1994.tb00397.x. [DOI] [PubMed] [Google Scholar]
  29. Modrich P. Mechanisms and biological effects of mismatch repair. Annu Rev Genet. 1991;25:229–253. doi: 10.1146/annurev.ge.25.120191.001305. [DOI] [PubMed] [Google Scholar]
  30. Moore M. D., Kaplan S. Identification of intrinsic high-level resistance to rare-earth oxides and oxyanions in members of the class Proteobacteria: characterization of tellurite, selenite, and rhodium sesquioxide reduction in Rhodobacter sphaeroides. J Bacteriol. 1992 Mar;174(5):1505–1514. doi: 10.1128/jb.174.5.1505-1514.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murphy R. C., Gasparich G. E., Bryant D. A., Porter R. D. Nucleotide sequence and further characterization of the Synechococcus sp. strain PCC 7002 recA gene: complementation of a cyanobacterial recA mutation by the Escherichia coli recA gene. J Bacteriol. 1990 Feb;172(2):967–976. doi: 10.1128/jb.172.2.967-976.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Perkins J. D., Heath J. D., Sharma B. R., Weinstock G. M. XbaI and BlnI genomic cleavage maps of Escherichia coli K-12 strain MG1655 and comparative analysis of other strains. J Mol Biol. 1993 Jul 20;232(2):419–445. doi: 10.1006/jmbi.1993.1401. [DOI] [PubMed] [Google Scholar]
  33. Sancar A., Sancar G. B. DNA repair enzymes. Annu Rev Biochem. 1988;57:29–67. doi: 10.1146/annurev.bi.57.070188.000333. [DOI] [PubMed] [Google Scholar]
  34. Sasakawa C., Yoshikawa M. A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis. Gene. 1987;56(2-3):283–288. doi: 10.1016/0378-1119(87)90145-4. [DOI] [PubMed] [Google Scholar]
  35. Sedgwick S. G., Goodwin P. A. Differences in mutagenic and recombinational DNA repair in enterobacteria. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4172–4176. doi: 10.1073/pnas.82.12.4172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Setlow R. B., Carrier W. L. Pyrimidine dimers in ultraviolet-irradiated DNA's. J Mol Biol. 1966 May;17(1):237–254. doi: 10.1016/s0022-2836(66)80105-5. [DOI] [PubMed] [Google Scholar]
  37. Sicard N. Possible correlation between transformability and deficiency in error-prone repair. J Bacteriol. 1983 May;154(2):995–997. doi: 10.1128/jb.154.2.995-997.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Smith D. W., Yee T. W., Baird C., Krishnapillai V. Pseudomonad replication origins: a paradigm for bacterial origins? Mol Microbiol. 1991 Nov;5(11):2581–2587. doi: 10.1111/j.1365-2958.1991.tb01966.x. [DOI] [PubMed] [Google Scholar]
  39. Stamm L. V., Charon N. W. Sensitivity of pathogenic and free-living Leptospira spp. to UV radiation and mitomycin C. Appl Environ Microbiol. 1988 Mar;54(3):728–733. doi: 10.1128/aem.54.3.728-733.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Steele M. I., Lorenz D., Hatter K., Park A., Sokatch J. R. Characterization of the mmsAB operon of Pseudomonas aeruginosa PAO encoding methylmalonate-semialdehyde dehydrogenase and 3-hydroxyisobutyrate dehydrogenase. J Biol Chem. 1992 Jul 5;267(19):13585–13592. [PubMed] [Google Scholar]
  41. Suwanto A., Kaplan S. Physical and genetic mapping of the Rhodobacter sphaeroides 2.4.1 genome: genome size, fragment identification, and gene localization. J Bacteriol. 1989 Nov;171(11):5840–5849. doi: 10.1128/jb.171.11.5840-5849.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Suwanto A., Kaplan S. Physical and genetic mapping of the Rhodobacter sphaeroides 2.4.1 genome: presence of two unique circular chromosomes. J Bacteriol. 1989 Nov;171(11):5850–5859. doi: 10.1128/jb.171.11.5850-5859.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tessman I., Liu S. K., Kennedy M. A. Mechanism of SOS mutagenesis of UV-irradiated DNA: mostly error-free processing of deaminated cytosine. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1159–1163. doi: 10.1073/pnas.89.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Unrau P., Wheatcroft R., Cox B., Olive T. The formation of pyrimidine dimers in the DNA of fungi and bacteria. Biochim Biophys Acta. 1973 Jul 27;312(4):626–632. doi: 10.1016/0005-2787(73)90065-8. [DOI] [PubMed] [Google Scholar]
  45. Van Houten B. Nucleotide excision repair in Escherichia coli. Microbiol Rev. 1990 Mar;54(1):18–51. doi: 10.1128/mr.54.1.18-51.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vericat J. A., Barbé J. Adaptive response to simple alkylating agents in the phototrophic bacteria Rhodobacter capsulatus and R.sphaeroides. Mutagenesis. 1988 Mar;3(2):165–168. doi: 10.1093/mutage/3.2.165. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES