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. 1995 Jan;177(1):86–93. doi: 10.1128/jb.177.1.86-93.1995

The rec locus, a competence-induced operon in Streptococcus pneumoniae.

B J Pearce 1, A M Naughton 1, E A Campbell 1, H R Masure 1
PMCID: PMC176560  PMID: 7798154

Abstract

To study competence and the process of transformation (TFN) in pneumococci, we developed a method for isolating TFN- mutants using insertional inactivation coupled with fusions to the gene for alkaline phosphatase (phoA). One TFN- mutant transformed 2 log units less efficiently than the parent strain. Reconstitution of the mutated region revealed a locus, rec, that contains two polycistronic genes, exp10 and the previously identified recA (B. Martin, J. M. Ruellan, J. F. Angulo, R. Devoret, and J. P. Claverys, Nucleic Acids Res. 20:6412, 1992). Exp10 is likely to be a membrane-associated protein, as it has a prokaryotic signal sequence and an Exp10-PhoA fusion localized with cell membranes. On the basis of sequence similarity, pneumococcal RecA is a member of bacterial RecA proteins responsible for homologous recombination of DNA. DNA-RNA hybridization analysis showed that this locus is transcribed as a polycistronic message, with increased transcription occurring during competence. With an Exp10-PhoA chimera used as a reporter, there was a 10-fold increase in the expression of the rec locus during competence while there was only minimal expression under growth conditions that repressed competence. The TFN- mutant containing the exp10-phoA fusion produced activator, a small extracellular polypeptide that induces competence, and the expression of rec was induced in response to activator. Therefore, the rec locus is directly required for genetic transformation and is regulated by the cell signaling mechanism that induces competence.

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Selected References

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  1. Ariza R. R., Cohen S. P., Bachhawat N., Levy S. B., Demple B. Repressor mutations in the marRAB operon that activate oxidative stress genes and multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1994 Jan;176(1):143–148. doi: 10.1128/jb.176.1.143-148.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen J. D., Morrison D. A. Construction and properties of a new insertion vector, pJDC9, that is protected by transcriptional terminators and useful for cloning of DNA from Streptococcus pneumoniae. Gene. 1988 Apr 15;64(1):155–164. doi: 10.1016/0378-1119(88)90489-1. [DOI] [PubMed] [Google Scholar]
  3. Cheo D. L., Bayles K. W., Yasbin R. E. Cloning and characterization of DNA damage-inducible promoter regions from Bacillus subtilis. J Bacteriol. 1991 Mar;173(5):1696–1703. doi: 10.1128/jb.173.5.1696-1703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheo D. L., Bayles K. W., Yasbin R. E. Elucidation of regulatory elements that control damage induction and competence induction of the Bacillus subtilis SOS system. J Bacteriol. 1993 Sep;175(18):5907–5915. doi: 10.1128/jb.175.18.5907-5915.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheo D. L., Bayles K. W., Yasbin R. E. Molecular characterization of regulatory elements controlling expression of the Bacillus subtilis recA+ gene. Biochimie. 1992 Jul-Aug;74(7-8):755–762. doi: 10.1016/0300-9084(92)90148-8. [DOI] [PubMed] [Google Scholar]
  6. Cohen S. P., Hächler H., Levy S. B. Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1484–1492. doi: 10.1128/jb.175.5.1484-1492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen S. P., Levy S. B., Foulds J., Rosner J. L. Salicylate induction of antibiotic resistance in Escherichia coli: activation of the mar operon and a mar-independent pathway. J Bacteriol. 1993 Dec;175(24):7856–7862. doi: 10.1128/jb.175.24.7856-7862.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. De Mot R., Schoofs G., Vanderleyden J. A putative regulatory gene downstream of recA is conserved in gram-negative and gram-positive bacteria. Nucleic Acids Res. 1994 Apr 11;22(7):1313–1314. doi: 10.1093/nar/22.7.1313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dubnau D. Genetic competence in Bacillus subtilis. Microbiol Rev. 1991 Sep;55(3):395–424. doi: 10.1128/mr.55.3.395-424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fyfe J. A., Davies J. K. Nucleotide sequence and expression in Escherichia coli of the recA gene of Neisseria gonorrhoeae. Gene. 1990 Sep 1;93(1):151–156. doi: 10.1016/0378-1119(90)90151-g. [DOI] [PubMed] [Google Scholar]
  12. Guenzi E., Gasc A. M., Sicard M. A., Hakenbeck R. A two-component signal-transducing system is involved in competence and penicillin susceptibility in laboratory mutants of Streptococcus pneumoniae. Mol Microbiol. 1994 May;12(3):505–515. doi: 10.1111/j.1365-2958.1994.tb01038.x. [DOI] [PubMed] [Google Scholar]
  13. Hakenbeck R., Ellerbrok H., Briese T., Handwerger S., Tomasz A. Penicillin-binding proteins of penicillin-susceptible and -resistant pneumococci: immunological relatedness of altered proteins and changes in peptides carrying the beta-lactam binding site. Antimicrob Agents Chemother. 1986 Oct;30(4):553–558. doi: 10.1128/aac.30.4.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hui F. M., Morrison D. A. Genetic transformation in Streptococcus pneumoniae: nucleotide sequence analysis shows comA, a gene required for competence induction, to be a member of the bacterial ATP-dependent transport protein family. J Bacteriol. 1991 Jan;173(1):372–381. doi: 10.1128/jb.173.1.372-381.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kirsch J., Wolters I., Triller A., Betz H. Gephyrin antisense oligonucleotides prevent glycine receptor clustering in spinal neurons. Nature. 1993 Dec 23;366(6457):745–748. doi: 10.1038/366745a0. [DOI] [PubMed] [Google Scholar]
  16. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  17. Lazarevic V., Margot P., Soldo B., Karamata D. Sequencing and analysis of the Bacillus subtilis lytRABC divergon: a regulatory unit encompassing the structural genes of the N-acetylmuramoyl-L-alanine amidase and its modifier. J Gen Microbiol. 1992 Sep;138(9):1949–1961. doi: 10.1099/00221287-138-9-1949. [DOI] [PubMed] [Google Scholar]
  18. Lovett C. M., Jr, Love P. E., Yasbin R. E. Competence-specific induction of the Bacillus subtilis RecA protein analog: evidence for dual regulation of a recombination protein. J Bacteriol. 1989 May;171(5):2318–2322. doi: 10.1128/jb.171.5.2318-2322.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Martin B., Humbert O., Camara M., Guenzi E., Walker J., Mitchell T., Andrew P., Prudhomme M., Alloing G., Hakenbeck R. A highly conserved repeated DNA element located in the chromosome of Streptococcus pneumoniae. Nucleic Acids Res. 1992 Jul 11;20(13):3479–3483. doi: 10.1093/nar/20.13.3479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Martin B., Ruellan J. M., Angulo J. F., Devoret R., Claverys J. P. Identification of the recA gene of Streptococcus pneumoniae. Nucleic Acids Res. 1992 Dec 11;20(23):6412–6412. doi: 10.1093/nar/20.23.6412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morrison D. A., Baker M. F. Competence for genetic transformation in pneumococcus depends on synthesis of a small set of proteins. Nature. 1979 Nov 8;282(5735):215–217. doi: 10.1038/282215a0. [DOI] [PubMed] [Google Scholar]
  23. Morrison D. A., Lacks S. A., Guild W. R., Hageman J. M. Isolation and characterization of three new classes of transformation-deficient mutants of Streptococcus pneumoniae that are defective in DNA transport and genetic recombination. J Bacteriol. 1983 Oct;156(1):281–290. doi: 10.1128/jb.156.1.281-290.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morrison D. A., Mannarelli B. Transformation in pneumococcus: nuclease resistance of deoxyribonucleic acid in the eclipse complex. J Bacteriol. 1979 Nov;140(2):655–665. doi: 10.1128/jb.140.2.655-665.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morrison D. A. Transformation in pneumococcus: existence and properties of a complex involving donor deoxyribonucleate single strands in eclipse. J Bacteriol. 1977 Nov;132(2):576–583. doi: 10.1128/jb.132.2.576-583.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morrison D. A. Transformation in pneumococcus: protein content of eclipse complex. J Bacteriol. 1978 Nov;136(2):548–557. doi: 10.1128/jb.136.2.548-557.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morrison D. A., Trombe M. C., Hayden M. K., Waszak G. A., Chen J. D. Isolation of transformation-deficient Streptococcus pneumoniae mutants defective in control of competence, using insertion-duplication mutagenesis with the erythromycin resistance determinant of pAM beta 1. J Bacteriol. 1984 Sep;159(3):870–876. doi: 10.1128/jb.159.3.870-876.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nohno T., Kasai Y., Saito T. Cloning and sequencing of the Escherichia coli chlEN operon involved in molybdopterin biosynthesis. J Bacteriol. 1988 Sep;170(9):4097–4102. doi: 10.1128/jb.170.9.4097-4102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pearce B. J., Naughton A. M., Masure H. R. Peptide permeases modulate transformation in Streptococcus pneumoniae. Mol Microbiol. 1994 Jun;12(6):881–892. doi: 10.1111/j.1365-2958.1994.tb01076.x. [DOI] [PubMed] [Google Scholar]
  30. Pearce B. J., Yin Y. B., Masure H. R. Genetic identification of exported proteins in Streptococcus pneumoniae. Mol Microbiol. 1993 Sep;9(5):1037–1050. doi: 10.1111/j.1365-2958.1993.tb01233.x. [DOI] [PubMed] [Google Scholar]
  31. Prior P., Schmitt B., Grenningloh G., Pribilla I., Multhaup G., Beyreuther K., Maulet Y., Werner P., Langosch D., Kirsch J. Primary structure and alternative splice variants of gephyrin, a putative glycine receptor-tubulin linker protein. Neuron. 1992 Jun;8(6):1161–1170. doi: 10.1016/0896-6273(92)90136-2. [DOI] [PubMed] [Google Scholar]
  32. Puyet A., Greenberg B., Lacks S. A. Genetic and structural characterization of endA. A membrane-bound nuclease required for transformation of Streptococcus pneumoniae. J Mol Biol. 1990 Jun 20;213(4):727–738. doi: 10.1016/S0022-2836(05)80259-1. [DOI] [PubMed] [Google Scholar]
  33. Puyet A., Ibáez A. M., Espinosa M. Characterization of the Streptococcus pneumoniae maltosaccharide regulator MalR, a member of the LacI-GalR family of repressors displaying distinctive genetic features. J Biol Chem. 1993 Dec 5;268(34):25402–25408. [PubMed] [Google Scholar]
  34. Radnis B. A., Rhee D. K., Morrison D. A. Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP. J Bacteriol. 1990 Jul;172(7):3669–3674. doi: 10.1128/jb.172.7.3669-3674.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Raymond-Denise A., Guillen N. Expression of the Bacillus subtilis dinR and recA genes after DNA damage and during competence. J Bacteriol. 1992 May;174(10):3171–3176. doi: 10.1128/jb.174.10.3171-3176.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Reizer J., Reizer A., Bairoch A., Saier M. H., Jr A diverse transketolase family that includes the RecP protein of Streptococcus pneumoniae, a protein implicated in genetic recombination. Res Microbiol. 1993 Jun;144(5):341–347. doi: 10.1016/0923-2508(93)90191-4. [DOI] [PubMed] [Google Scholar]
  37. Rhee D. K., Morrison D. A. Genetic transformation in Streptococcus pneumoniae: molecular cloning and characterization of recP, a gene required for genetic recombination. J Bacteriol. 1988 Feb;170(2):630–637. doi: 10.1128/jb.170.2.630-637.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tiraby J. G., Fox M. S. Marker discrimination in transformation and mutation of pneumococcus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3541–3545. doi: 10.1073/pnas.70.12.3541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tomasz A. Cellular metabolism in genetic transformation of pneumococci: requirement for protein synthesis during induction of competence. J Bacteriol. 1970 Mar;101(3):860–871. doi: 10.1128/jb.101.3.860-871.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tomasz A. Control of the competent state in Pneumococcus by a hormone-like cell product: an example for a new type of regulatory mechanism in bacteria. Nature. 1965 Oct 9;208(5006):155–159. doi: 10.1038/208155a0. [DOI] [PubMed] [Google Scholar]
  42. Tomasz A. Model for the mechanism controlling the expression of competent state in Pneumococcus cultures. J Bacteriol. 1966 Mar;91(3):1050–1061. doi: 10.1128/jb.91.3.1050-1061.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tomasz A., Mosser J. L. On the nature of the pneumococcal activator substance. Proc Natl Acad Sci U S A. 1966 Jan;55(1):58–66. doi: 10.1073/pnas.55.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vijayakumar M. N., Morrison D. A. Fate of DNA in eclipse complex during genetic transformation in Streptococcus pneumoniae. J Bacteriol. 1983 Nov;156(2):644–648. doi: 10.1128/jb.156.2.644-648.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zulty J. J., Barcak G. J. Structural organization, nucleotide sequence, and regulation of the Haemophilus influenzae rec-1+ gene. J Bacteriol. 1993 Nov;175(22):7269–7281. doi: 10.1128/jb.175.22.7269-7281.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. te Riele H. P., Venema G. Heterospecific transformation in Bacillus subtilis: protein composition of a membrane-DNA complex containing unstable heterologous donor-recipient complex. Mol Gen Genet. 1984;197(3):478–485. doi: 10.1007/BF00329946. [DOI] [PubMed] [Google Scholar]
  47. te Riele H. P., Venema G. Molecular fate of heterologous bacterial DNA in competent Bacillus subtilis: further characterization of unstable association between donor and recipient DNA and the involvement of the cellular membrane. Mol Gen Genet. 1984;195(1-2):200–208. doi: 10.1007/BF00332747. [DOI] [PubMed] [Google Scholar]
  48. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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