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. 1997 Apr;179(8):2783–2787. doi: 10.1128/jb.179.8.2783-2787.1997

Cloning and characterization of the ponA gene encoding penicillin-binding protein 1 from Neisseria gonorrhoeae and Neisseria meningitidis.

P A Ropp 1, R A Nicholas 1
PMCID: PMC179034  PMID: 9098083

Abstract

The ponA gene encoding penicillin-binding protein 1 (PBP 1) from Neisseria gonorrhoeae was cloned by a reverse genetic approach. PBP 1 was purified from solubilized membranes of penicillin-susceptible strain FA19 by covalent ampicillin affinity chromatography and used to obtain an NH2-terminal amino acid sequence. A degenerate oligonucleotide based on this protein sequence and a highly degenerate oligonucleotide based on a conserved amino acid motif found in all class A high-molecular-mass PBPs were used to isolate the PBP 1 gene (ponA). The ponA gene encodes a protein containing all of the conserved sequence motifs found in class A PBPs, and expression of the gene in Escherichia coli resulted in the appearance of a new PBP that comigrated with PBP 1 purified from N. gonorrhoeae. A comparison of the gonococcal ponA gene to its homolog isolated from Neisseria meningitidis revealed a high degree of identity between the two gene products, with the greatest variability found at the carboxy terminus of the two deduced PBP 1 protein sequences.

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Selected References

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  1. Barbour A. G. Properties of penicillin-binding proteins in Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1981 Feb;19(2):316–322. doi: 10.1128/aac.19.2.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Black C. G., Fyfe J. A., Davies J. K. A promoter associated with the neisserial repeat can be used to transcribe the uvrB gene from Neisseria gonorrhoeae. J Bacteriol. 1995 Apr;177(8):1952–1958. doi: 10.1128/jb.177.8.1952-1958.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cannon J. G., Sparling P. F. The genetics of the gonococcus. Annu Rev Microbiol. 1984;38:111–133. doi: 10.1146/annurev.mi.38.100184.000551. [DOI] [PubMed] [Google Scholar]
  4. Cornelissen C. N., Biswas G. D., Tsai J., Paruchuri D. K., Thompson S. A., Sparling P. F. Gonococcal transferrin-binding protein 1 is required for transferrin utilization and is homologous to TonB-dependent outer membrane receptors. J Bacteriol. 1992 Sep;174(18):5788–5797. doi: 10.1128/jb.174.18.5788-5797.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dideberg O., Charlier P., Wéry J. P., Dehottay P., Dusart J., Erpicum T., Frère J. M., Ghuysen J. M. The crystal structure of the beta-lactamase of Streptomyces albus G at 0.3 nm resolution. Biochem J. 1987 Aug 1;245(3):911–913. doi: 10.1042/bj2450911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dougherty T. J. Genetic analysis and penicillin-binding protein alterations in Neisseria gonorrhoeae with chromosomally mediated resistance. Antimicrob Agents Chemother. 1986 Nov;30(5):649–652. doi: 10.1128/aac.30.5.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dougherty T. J., Koller A. E., Tomasz A. Penicillin-binding proteins of penicillin-susceptible and intrinsically resistant Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1980 Nov;18(5):730–737. doi: 10.1128/aac.18.5.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Drake S. L., Koomey M. The product of the pilQ gene is essential for the biogenesis of type IV pili in Neisseria gonorrhoeae. Mol Microbiol. 1995 Dec;18(5):975–986. doi: 10.1111/j.1365-2958.1995.18050975.x. [DOI] [PubMed] [Google Scholar]
  9. Elkins C., Thomas C. E., Seifert H. S., Sparling P. F. Species-specific uptake of DNA by gonococci is mediated by a 10-base-pair sequence. J Bacteriol. 1991 Jun;173(12):3911–3913. doi: 10.1128/jb.173.12.3911-3913.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faruki H., Sparling P. F. Genetics of resistance in a non-beta-lactamase-producing gonococcus with relatively high-level penicillin resistance. Antimicrob Agents Chemother. 1986 Dec;30(6):856–860. doi: 10.1128/aac.30.6.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fleischmann R. D., Adams M. D., White O., Clayton R. A., Kirkness E. F., Kerlavage A. R., Bult C. J., Tomb J. F., Dougherty B. A., Merrick J. M. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995 Jul 28;269(5223):496–512. doi: 10.1126/science.7542800. [DOI] [PubMed] [Google Scholar]
  12. Fraser C. M., Gocayne J. D., White O., Adams M. D., Clayton R. A., Fleischmann R. D., Bult C. J., Kerlavage A. R., Sutton G., Kelley J. M. The minimal gene complement of Mycoplasma genitalium. Science. 1995 Oct 20;270(5235):397–403. doi: 10.1126/science.270.5235.397. [DOI] [PubMed] [Google Scholar]
  13. Ghuysen J. M., Frère J. M., Leyh-Bouille M., Nguyen-Distèche M., Coyette J. Active-site-serine D-alanyl-D-alanine-cleaving-peptidase-catalysed acyl-transfer reactions. Procedures for studying the penicillin-binding proteins of bacterial plasma membranes. Biochem J. 1986 Apr 1;235(1):159–165. doi: 10.1042/bj2350159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Herzberg O., Moult J. Bacterial resistance to beta-lactam antibiotics: crystal structure of beta-lactamase from Staphylococcus aureus PC1 at 2.5 A resolution. Science. 1987 May 8;236(4802):694–701. doi: 10.1126/science.3107125. [DOI] [PubMed] [Google Scholar]
  15. Kelly J. A., Knox J. R., Moews P. C., Hite G. J., Bartolone J. B., Zhao H., Joris B., Frère J. M., Ghuysen J. M. 2.8-A Structure of penicillin-sensitive D-alanyl carboxypeptidase-transpeptidase from Streptomyces R61 and complexes with beta-lactams. J Biol Chem. 1985 May 25;260(10):6449–6458. [PubMed] [Google Scholar]
  16. Martin P. R., Watson A. A., McCaul T. F., Mattick J. S. Characterization of a five-gene cluster required for the biogenesis of type 4 fimbriae in Pseudomonas aeruginosa. Mol Microbiol. 1995 May;16(3):497–508. doi: 10.1111/j.1365-2958.1995.tb02414.x. [DOI] [PubMed] [Google Scholar]
  17. Matsuhashi M., Maruyama I. N., Takagaki Y., Tamaki S., Nishimura Y., Hirota Y. Isolation of a mutant of Escherichia coli lacking penicillin-sensitive D-alanine carboxypeptidase IA. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2631–2635. doi: 10.1073/pnas.75.6.2631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moews P. C., Knox J. R., Dideberg O., Charlier P., Frère J. M. Beta-lactamase of Bacillus licheniformis 749/C at 2 A resolution. Proteins. 1990;7(2):156–171. doi: 10.1002/prot.340070205. [DOI] [PubMed] [Google Scholar]
  19. Pares S., Mouz N., Pétillot Y., Hakenbeck R., Dideberg O. X-ray structure of Streptococcus pneumoniae PBP2x, a primary penicillin target enzyme. Nat Struct Biol. 1996 Mar;3(3):284–289. doi: 10.1038/nsb0396-284. [DOI] [PubMed] [Google Scholar]
  20. Plunkett G., 3rd, Burland V., Daniels D. L., Blattner F. R. Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes. Nucleic Acids Res. 1993 Jul 25;21(15):3391–3398. doi: 10.1093/nar/21.15.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Popham D. L., Setlow P. Cloning, nucleotide sequence, mutagenesis, and mapping of the Bacillus subtilis pbpD gene, which codes for penicillin-binding protein 4. J Bacteriol. 1994 Dec;176(23):7197–7205. doi: 10.1128/jb.176.23.7197-7205.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roine E., Nunn D. N., Paulin L., Romantschuk M. Characterization of genes required for pilus expression in Pseudomonas syringae pathovar phaseolicola. J Bacteriol. 1996 Jan;178(2):410–417. doi: 10.1128/jb.178.2.410-417.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Suzuki H., Nishimura Y., Hirota Y. On the process of cellular division in Escherichia coli: a series of mutants of E. coli altered in the penicillin-binding proteins. Proc Natl Acad Sci U S A. 1978 Feb;75(2):664–668. doi: 10.1073/pnas.75.2.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tamura T., Suzuki H., Nishimura Y., Mizoguchi J., Hirota Y. On the process of cellular division in Escherichia coli: isolation and characterization of penicillin-binding proteins 1a, 1b, and 3. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4499–4503. doi: 10.1073/pnas.77.8.4499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tipper D. J., Strominger J. L. Mechanism of action of penicillins: a proposal based on their structural similarity to acyl-D-alanyl-D-alanine. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1133–1141. doi: 10.1073/pnas.54.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]

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