Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Aug;174(15):4899–4906. doi: 10.1128/jb.174.15.4899-4906.1992

Construction of a Neisseria gonorrhoeae MS11 derivative deficient in NgoMI restriction and modification.

D C Stein 1, R Chien 1, H S Seifert 1
PMCID: PMC206301  PMID: 1321116

Abstract

We have cloned from Neisseria gonorrhoeae MS11 the gene encoding a methylase that modifies the sequence GCCGGC. The corresponding restriction enzyme was also encoded by this clone. Sequence analysis demonstrated that the methylase shares sequence similarities with other cytosine methylases, but the sequence organization of M.NgoMI is different from that seen for other cytosine methylases. A deletion was introduced into the chromosome of N. gonorrhoeae MS11 to produce strain MUG701, a strain that is inactivated in both the methylase and the restriction genes. Although this strain no longer methylated its DNA at the NgoMI recognition sequence, cells were viable and had no other significant phenotypic changes. Transformation data indicated that MS11 does not produce enough restriction activity to block plasmid transformation in the gonococcus, even though restriction activity could be demonstrated in E. coli containing the cloned gene.

Full text

PDF
4901

Selected References

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

  1. ARBER W., MORSE M. L. HOST SPECIFICITY OF DNA PRODUCED BY ESCHERICHIA COLI. VI. EFFECTS ON BACTERIAL CONJUGATION. Genetics. 1965 Jan;51:137–148. doi: 10.1093/genetics/51.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Butler C. A., Gotschlich E. C. High-frequency mobilization of broad-host-range plasmids into Neisseria gonorrhoeae requires methylation in the donor. J Bacteriol. 1991 Sep;173(18):5793–5799. doi: 10.1128/jb.173.18.5793-5799.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clanton D. J., Woodward J. M., Miller R. V. Identification of a new sequence-specific endonuclease, NgoII, from Neisseria gonorrhoeae. J Bacteriol. 1978 Jul;135(1):270–273. doi: 10.1128/jb.135.1.270-273.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Conrad M., Topal M. D. DNA and spermidine provide a switch mechanism to regulate the activity of restriction enzyme Nae I. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9707–9711. doi: 10.1073/pnas.86.24.9707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DUSSOIX D., ARBER W. Host specificity of DNA produced by Escherichia coli. II. Control over acceptance of DNA from infecting phage lambda. J Mol Biol. 1962 Jul;5:37–49. doi: 10.1016/s0022-2836(62)80059-x. [DOI] [PubMed] [Google Scholar]
  6. Duff M. K., Davies J. K. Multiple restriction-modification systems in Neisseria gonorrhoeae. Gene. 1988 Dec 25;74(1):227–228. doi: 10.1016/0378-1119(88)90292-2. [DOI] [PubMed] [Google Scholar]
  7. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Korch C., Hagblom P., Normark S. Sequence-specific DNA modification in Neisseria gonorrhoeae. J Bacteriol. 1983 Sep;155(3):1324–1332. doi: 10.1128/jb.155.3.1324-1332.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Korch C., Hagblom P., Normark S. Type III 5-methylcytosine modification of DNA in Neisseria gonorrhoeae. J Bacteriol. 1985 Mar;161(3):1236–1237. doi: 10.1128/jb.161.3.1236-1237.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Krüger D. H., Bickle T. A. Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts. Microbiol Rev. 1983 Sep;47(3):345–360. doi: 10.1128/mr.47.3.345-360.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lauster R. Evolution of type II DNA methyltransferases. A gene duplication model. J Mol Biol. 1989 Mar 20;206(2):313–321. doi: 10.1016/0022-2836(89)90481-6. [DOI] [PubMed] [Google Scholar]
  12. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  13. Moseley S. L., Huq I., Alim A. R., So M., Samadpour-Motalebi M., Falkow S. Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis. 1980 Dec;142(6):892–898. doi: 10.1093/infdis/142.6.892. [DOI] [PubMed] [Google Scholar]
  14. Nathan P. D., Brooks J. E. Characterization of clones of the BamHI methyltransferase gene. Gene. 1988 Dec 25;74(1):35–36. doi: 10.1016/0378-1119(88)90244-2. [DOI] [PubMed] [Google Scholar]
  15. Norlander L., Davies J. K., Hagblom P., Normark S. Deoxyribonucleic acid modifications and restriction endonuclease production in Neisseria gonorrhoeae. J Bacteriol. 1981 Feb;145(2):788–795. doi: 10.1128/jb.145.2.788-795.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Piekarowicz A., Yuan R., Stein D. C. Identification of a new restriction endonuclease, R.NgoBI, from Neisseria gonorrhoeae. Nucleic Acids Res. 1988 Oct 25;16(20):9868–9868. doi: 10.1093/nar/16.20.9868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Piekarowicz A., Yuan R., Stein D. C. Purification and characterization of DNA methyltransferases from Neisseria gonorrhoeae. Nucleic Acids Res. 1988 Jul 11;16(13):5957–5972. doi: 10.1093/nar/16.13.5957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pridmore R. D. New and versatile cloning vectors with kanamycin-resistance marker. Gene. 1987;56(2-3):309–312. doi: 10.1016/0378-1119(87)90149-1. [DOI] [PubMed] [Google Scholar]
  19. Roberts R. J. Restriction enzymes and their isoschizomers. Nucleic Acids Res. 1988;16 (Suppl):r271–r313. doi: 10.1093/nar/16.suppl.r271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  21. Segal E., Hagblom P., Seifert H. S., So M. Antigenic variation of gonococcal pilus involves assembly of separated silent gene segments. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2177–2181. doi: 10.1073/pnas.83.7.2177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Seifert H. S., Chen E. Y., So M., Heffron F. Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1986 Feb;83(3):735–739. doi: 10.1073/pnas.83.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stein D. C., Danaher R. J., Cook T. M. Characterization of a gyrB mutation responsible for low-level nalidixic acid resistance in Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1991 Apr;35(4):622–626. doi: 10.1128/aac.35.4.622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stein D. C., Gregoire S., Piekarowicz A. Restriction of plasmid DNA during transformation but not conjugation in Neisseria gonorrhoeae. Infect Immun. 1988 Jan;56(1):112–116. doi: 10.1128/iai.56.1.112-116.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stein D. C. Introduction of cloned genes into Neisseria gonorrhoeae. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S146–S149. doi: 10.1128/cmr.2.suppl.s146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stein D. C. Transformation of Neisseria gonorrhoeae: physical requirements of the transforming DNA. Can J Microbiol. 1991 May;37(5):345–349. doi: 10.1139/m91-056. [DOI] [PubMed] [Google Scholar]
  27. Stein D. C., Young F. E., Tenover F. C., Clark V. L. Characterization of a chimeric beta-lactamase plasmid of Neisseria gonorrhoeae which can function in Escherichia coli. Mol Gen Genet. 1983;189(1):77–84. doi: 10.1007/BF00326058. [DOI] [PubMed] [Google Scholar]
  28. Sullivan K. M., Saunders J. R. Nucleotide sequence and genetic organization of the NgoPII restriction-modification system of Neisseria gonorrhoeae. Mol Gen Genet. 1989 Apr;216(2-3):380–387. doi: 10.1007/BF00334379. [DOI] [PubMed] [Google Scholar]
  29. Taha M. K., So M., Seifert H. S., Billyard E., Marchal C. Pilin expression in Neisseria gonorrhoeae is under both positive and negative transcriptional control. EMBO J. 1988 Dec 20;7(13):4367–4378. doi: 10.1002/j.1460-2075.1988.tb03335.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wu J. C., Santi D. V. Kinetic and catalytic mechanism of HhaI methyltransferase. J Biol Chem. 1987 Apr 5;262(10):4778–4786. [PubMed] [Google Scholar]

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

RESOURCES