PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
 
J Bacteriol. 1991 September; 173(18): 5793–5799.
PMCID: PMC208312

High-frequency mobilization of broad-host-range plasmids into Neisseria gonorrhoeae requires methylation in the donor.

Abstract

Antibiotic resistance in Neisseria gonorrhoeae has been associated with the acquisition of R plasmids from heterologous organisms. The broad-host-range plasmids of incompatibility groups P (IncP) and Q (IncQ) have played a role in this genetic exchange in nature. We have utilized derivatives of RSF1010 (IncQ) and RP1 (IncP) to demonstrate that the plethora of restriction barriers associated with the gonococci markedly reduces mobilization of plasmids from Escherichia coli into strains F62 and PGH 3-2. Partially purified restriction endonucleases from these gonococcal strains can digest RSF1010 in vitro. Protection of RSF1010-km from digestion by gonococcal enzymes purified from strain F62 is observed when the plasmid is isolated from E. coli containing a coresident plasmid, pCAL7. Plasmid pCAL7 produces a 5'-MECG-3' cytosine methylase (M.SssI). The M.SssI methylase only partially protects RSF1010-km from digestion by restriction enzymes from strain PGH 3-2. Total protection of RSF1010-km from PGH 3-2 restriction requires both pCAL7 and a second coresident plasmid, pFnuDI, which produces a 5'-GGMECC-3' cytosine methylase. When both F62 and PGH 3-2 are utilized as recipients in heterospecific matings with E. coli, mobilization of RSF1010 from strains containing the appropriate methylases into the gonococci occurs at frequencies 4 orders of magnitude higher than from strains without the methylases. Thus, protection of RSF1010 from gonococcal restriction enzymes in vitro correlates with an increase in the conjugal frequency. These data indicate that restriction is a major barrier against efficient conjugal transfer between N. gonorrhoeae and heterologous hosts.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Burkardt HJ, Riess G, Pühler A. Relationship of group P1 plasmids revealed by heteroduplex experiments: RP1, RP4, R68 and RK2 are identical. J Gen Microbiol. 1979 Oct;114(2):341–348. [PubMed]
  • Davies JK. DNA restriction and modification systems in Neisseria gonorrhoeae. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S78–S82. [PMC free article] [PubMed]
  • Ditta G, Stanfield S, Corbin D, Helinski DR. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. [PubMed]
  • Facinelli B, Varaldo PE. Plasmid-mediated sulfonamide resistance in Neisseria meningitidis. Antimicrob Agents Chemother. 1987 Oct;31(10):1642–1643. [PMC free article] [PubMed]
  • Goodman SD, Scocca JJ. Identification and arrangement of the DNA sequence recognized in specific transformation of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6982–6986. [PubMed]
  • Graves JF, Biswas GD, Sparling PF. Sequence-specific DNA uptake in transformation of Neisseria gonorrhoeae. J Bacteriol. 1982 Dec;152(3):1071–1077. [PMC free article] [PubMed]
  • Guiney DG., Jr Promiscuous transfer of drug resistance in gram-negative bacteria. J Infect Dis. 1984 Mar;149(3):320–329. [PubMed]
  • Korch C, Hagblom P, Normark S. Sequence-specific DNA modification in Neisseria gonorrhoeae. J Bacteriol. 1983 Sep;155(3):1324–1332. [PMC free article] [PubMed]
  • Korch C, Hagblom P, Normark S. Type III 5-methylcytosine modification of DNA in Neisseria gonorrhoeae. J Bacteriol. 1985 Mar;161(3):1236–1237. [PMC free article] [PubMed]
  • Marra A, Shuman HA. Isolation of a Legionella pneumophila restriction mutant with increased ability to act as a recipient in heterospecific matings. J Bacteriol. 1989 Apr;171(4):2238–2240. [PMC free article] [PubMed]
  • Meinkoth J, Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. [PubMed]
  • Nur I, Szyf M, Razin A, Glaser G, Rottem S, Razin S. Procaryotic and eucaryotic traits of DNA methylation in spiroplasmas (mycoplasmas). J Bacteriol. 1985 Oct;164(1):19–24. [PMC free article] [PubMed]
  • Piffaretti JC, Arini A, Frey J. pUB307 mobilizes resistance plasmids from Escherichia coli into Neisseria gonorrhoeae. Mol Gen Genet. 1988 May;212(2):215–218. [PubMed]
  • Pintado C, Salvador C, Rotger R, Nombela C. Multiresistance plasmid from commensal Neisseria strains. Antimicrob Agents Chemother. 1985 Jan;27(1):120–124. [PMC free article] [PubMed]
  • Pirrotta V, Bickle TA. General purification schemes for restriction endonucleases. Methods Enzymol. 1980;65(1):89–95. [PubMed]
  • Renbaum P, Abrahamove D, Fainsod A, Wilson GG, Rottem S, Razin A. Cloning, characterization, and expression in Escherichia coli of the gene coding for the CpG DNA methylase from Spiroplasma sp. strain MQ1(M.SssI). Nucleic Acids Res. 1990 Mar 11;18(5):1145–1152. [PMC free article] [PubMed]
  • Roberts M, Falkow S. Conjugal transfer of R plasmids in Neisseria gonorrhoeae. Nature. 1977 Apr 14;266(5603):630–631. [PubMed]
  • Roberts MC. Plasmids of Neisseria gonorrhoeae and other Neisseria species. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S18–S23. [PMC free article] [PubMed]
  • Rotger R, Rubio F, Nombela C. A multi-resistance plasmid isolated from commensal Neisseria species is closely related to the enterobacterial plasmid RSF1010. J Gen Microbiol. 1986 Sep;132(9):2491–2496. [PubMed]
  • Schmidhauser TJ, Helinski DR. Regions of broad-host-range plasmid RK2 involved in replication and stable maintenance in nine species of gram-negative bacteria. J Bacteriol. 1985 Oct;164(1):446–455. [PMC free article] [PubMed]
  • Scholz P, Haring V, Wittmann-Liebold B, Ashman K, Bagdasarian M, Scherzinger E. Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010. Gene. 1989 Feb 20;75(2):271–288. [PubMed]
  • Sox TE, Mohammed W, Sparling PF. Transformation-derived Neisseria gonorrhoeae plasmids with altered structure and function. J Bacteriol. 1979 May;138(2):510–518. [PMC free article] [PubMed]
  • Stein DC, Gregoire S, Piekarowicz A. Restriction of plasmid DNA during transformation but not conjugation in Neisseria gonorrhoeae. Infect Immun. 1988 Jan;56(1):112–116. [PMC free article] [PubMed]
  • Swanson J. Studies on gonococcus infection. XIV. Cell wall protein differences among color/opacity colony variants of Neisseria gonorrhoeae. Infect Immun. 1978 Jul;21(1):292–302. [PMC free article] [PubMed]
  • Wells RD, Neuendorf SK. Cleavage of "single-stranded" viral DNAs by certain restriction endonucleases. Gene Amplif Anal. 1981;1:101–111. [PubMed]

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