1887

Abstract

Sulphonamide resistance in some clinical isolates of is associated with an insertion in the chromosomal gene leading to the addition of two amino acids, serine and glycine, in the drug target enzyme dihydropteroate synthase (DHPS). Removal of the insertion resulted in a markedly higher for the substrate -aminobenzoic acid and a markedly lower for 2-amino-4-hydroxy-6-(hydroxymethyl)-7,8-dihydropteridine pyrophosphate. In the same isolates an additional important difference, compared to wild-type enzymes, was found at amino acid position 68, which is a proline in most DHPS enzymes, but is serine in one and leucine in another clinical isolate of sulphonamide-resistant . The alteration at position 68 was found to affect mainly the level of sulphonamide resistance and had only a minor effect on the for the substrates. Introduction of the serine-glycine dipeptide at position 194 and a proline to serine substitution at position 68 in DHPS from normal, susceptible failed to produce a functional sulphonamide-resistant enzyme. The conclusion of this study is that it is not possibile to change a normal chromosomally encoded DHPS of to a sulphonamide-resistant one simply by an insertion of serine and glycine as seen in clinical isolates. It is likely that the resistance gene found in clinical isolates has evolved in another bacterial species where a combination of other amino acid changes may have contributed to produce a functionally resistant enzyme. This new resistance gene may have then been introduced into by natural transformation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-5-1151
2000-05-01
2019-10-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/5/1461151a.html?itemId=/content/journal/micro/10.1099/00221287-146-5-1151&mimeType=html&fmt=ahah

References

  1. Achari, A., Somers, D. O., Champness, J. N., Bryant, P. K., Rosemond, J. & Stammers, D. K. ( 1997; ). Crystal structure of the anti-bacterial sulphonamide drug target dihydropteroate synthase. Nat Struct Biol 4, 490-497.[CrossRef]
    [Google Scholar]
  2. Bertani, G. ( 1951; ). Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62, 293-300.
    [Google Scholar]
  3. Björkman, J., Hughes, D. & Andersson, D. I. ( 1998; ). Virulence of antibiotic-resistant Salmonella typhimurium. Proc Natl Acad Sci USA 95, 3949-3953.[CrossRef]
    [Google Scholar]
  4. Brooks, D. R., Wang, P., Read, M., Watkins, W. M., Sims, P. F. G. & Hyde, J. E. ( 1994; ). Sequence variation of the hydroxymethyldihydropterin pyrophosphokinase: dihydropteroate synthase gene in lines of the human malaria parasite, Plasmodium falciparum, with differing resistance to sulphadoxine. Eur J Biochem 224, 397-405.[CrossRef]
    [Google Scholar]
  5. Dallas, W. R., Gowen, J. E., Ray, P. H., Cox, M. J. & Dev, K. I. ( 1992; ). Cloning, sequencing and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100. J Bacteriol 174, 5961-5970.
    [Google Scholar]
  6. Dower, W. J., Miller, J. F. & Ragsdale, C. W. ( 1988; ). High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16, 6127-6145.[CrossRef]
    [Google Scholar]
  7. Fermér, C. & Swedberg, G. ( 1997; ). Adaptation to sulphonamide resistance in Neisseria meningitidis may have required compensatory changes to retain enzyme function: kinetic analysis of dihydropteroate synthases from N. meningitidis expressed in a knockout mutant of E. coli. J Bacteriol 179, 831-837.
    [Google Scholar]
  8. Fermér, C., Kristiansen, B.-E., Sköld, O. & Swedberg, G. ( 1995; ). Sulphonamide resistance in Neisseria meningitidis as defined by site-directed mutagenesis could have its origin in other species. J Bacteriol 177, 4669-4675.
    [Google Scholar]
  9. Hampele, I. C., D’Arcy, A., Dale, G. E. & 7 other authors ( 1997; ). Structure and function of the dihydropteroate synthase from Staphylococcus aureus. J Mol Biol 286, 21–30.
    [Google Scholar]
  10. Huovinen, P., Sundström, L., Swedberg, G. & Sköld, O. ( 1995; ). Trimethoprim and sulphonamide resistance. Antimicrob Agents Chemother 39, 279-289.[CrossRef]
    [Google Scholar]
  11. Levin, B. R., Lipsitch, M., Perrot, V., Schrag, S., Antia, R., Simonsen, L., Moore Walker, N. & Stewart, F. M. ( 1997; ). The population genetics of antibiotic resistance. Clin Infect Dis 24, S9-S16.[CrossRef]
    [Google Scholar]
  12. Lopez, P., Espinosa, M., Greenberg, B. & Lacks, S. A. ( 1987; ). Sulphonamide resistance in Streptococcus pneumoniae: DNA sequence of the gene encoding dihydropteroate synthase and characterization of the enzyme. J Bacteriol 169, 4320-4326.
    [Google Scholar]
  13. Maskell, J. P., Sefton, A. M. & Hall, L. M. C. ( 1997; ). Mechanism of sulphonamide resistance in clinical isolates of Streptococcus pneumoniae. Antimicrob Agents Chemother 41, 2121-2126.
    [Google Scholar]
  14. Müller-Hill, B., Crapo, L. & Gilbert, W. ( 1968; ). Mutants that make more lac repressor. Proc Natl Acad Sci USA 59, 1259-1264.[CrossRef]
    [Google Scholar]
  15. Rådström, P., Fermér, C., Kristiansen, B.-E., Jenkins, A., Sköld, O. & Swedberg, G. ( 1992; ). Transformational exchanges in the dihydropteroate synthase gene of Neisseria meningitidis: a novel mechanism for acquisition of sulphonamide resistance. J Bacteriol 174, 6386-6393.
    [Google Scholar]
  16. Sarkar, G. & Sommer, S. S. ( 1990; ). The ‘megaprimer’ method of site-directed mutagenesis Biotechniques 8, 404-407.
    [Google Scholar]
  17. Schrag, S. J. & Perrot, V. ( 1996; ). Reducing antibiotic resistance. Nature 381, 120-121.
    [Google Scholar]
  18. Spratt, B. G. ( 1994; ). Resistance to antibiotics mediated by target alterations. Science 264, 388-393.[CrossRef]
    [Google Scholar]
  19. Stuart, F. M., Antia, R., Levin, B. R., Lipsitch, M. & Mittler, J. E. ( 1998; ). The population genetics of antibiotic resistance II: analytic theory for sustained populations of bacteria in a community of hosts. Theor Popul Biol 53, 152-165.[CrossRef]
    [Google Scholar]
  20. Swedberg, G. & Sköld, O. ( 1980; ). Characterization of different plasmid-borne dihydropteroate synthases mediating bacterial resistance to sulphonamides. J Bacteriol 142, 1-7.
    [Google Scholar]
  21. Triglia, T. & Cowman, A. F. ( 1994; ). Primary structure and expression of the dihydropteroate synthase gene of Plasmodium falciparum. Proc Natl Acad Sci USA 91, 7141-7153.
    [Google Scholar]
  22. Triglia, T., Wang, P., Sims, P. F. G., Hyde, J. E. & Cowman, A. F. ( 1998; ). Allelic exchange at the endogenous genomic locus in Plasmodium falciparum proves the role of dihydropteroate synthase in sulphadoxine-resistant malaria. EMBO J 17, 3807-3815.[CrossRef]
    [Google Scholar]
  23. Vandeyar, M. A., Weiner, M. P., Hutton, C. J. & Batt, C. A. ( 1988; ). A simple and rapid method for the selection of oligodeoxynucleotide-directed mutants. Gene 65, 129-133.[CrossRef]
    [Google Scholar]
  24. Vedantam, G., Guay, G. G., Austria, N. E., Doktor, S. Z. & Nichols, B. P. ( 1998; ). Characterization of mutations contributing to sulphathiazole resistance in Escherichia coli. Antimicrob Agents Chemother 42, 88-93.
    [Google Scholar]
  25. Volpe, F., Dyer, M., Scaife, J. G., Darby, G., Stammers, D. K. & Delves, C. J. ( 1992; ). The multifunctional folic acid synthesis fas gene of Pneumocystis carinii appears to encode dihydropteroate synthase and hydroxymethyldihydropterin pyrophosphokinase. Gene 112, 213-218.[CrossRef]
    [Google Scholar]
  26. Yannisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103-119.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-5-1151
Loading
/content/journal/micro/10.1099/00221287-146-5-1151
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error