Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1994 Dec;38(12):2850–2856. doi: 10.1128/aac.38.12.2850

Characterization of the PNT1 pentamidine resistance gene of Saccharomyces cerevisiae.

G Ludewig 1, C Staben 1
PMCID: PMC188296  PMID: 7695273

Abstract

The Saccharomyces cerevisiae PNT1 gene was isolated and characterized. When present in high copy number in S. cerevisiae, PNT1 confers resistance to the anti-Pneumocystis carinii drug pentamidine. The PNT1 gene encodes a previously uncharacterized polypeptide of 409 amino acids. The predicted gene product is a very basic (pI 9.9) polypeptide with one potential membrane-associated region. PNT1 is located on chromosome XVR of S. cerevisiae. It is transcribed at a very low level. Overexpression of the gene increases resistance to the cytostatic and mitochondrial DNA-damaging effects of pentamidine and related cationic compounds. Disruption of the gene leads to slightly increased levels of susceptibility to pentamidine and some related compounds.

Full text

PDF
2850

Images in this article

2853Image
on p.2853
2853Image
on p.2853

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  3. Bruns T. D., Vilgalys R., Barns S. M., Gonzalez D., Hibbett D. S., Lane D. J., Simon L., Stickel S., Szaro T. M., Weisburg W. G. Evolutionary relationships within the fungi: analyses of nuclear small subunit rRNA sequences. Mol Phylogenet Evol. 1992 Sep;1(3):231–241. doi: 10.1016/1055-7903(92)90020-h. [DOI] [PubMed] [Google Scholar]
  4. Comley J. C., Mullin R. J., Wolfe L. A., Hanlon M. H., Ferone R. Microculture screening assay for primary in vitro evaluation of drugs against Pneumocystis carinii. Antimicrob Agents Chemother. 1991 Oct;35(10):1965–1974. doi: 10.1128/aac.35.10.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Damper D., Patton C. L. Pentamidine transport and sensitivity in brucei-group trypanosomes. J Protozool. 1976 May;23(2):349–356. doi: 10.1111/j.1550-7408.1976.tb03787.x. [DOI] [PubMed] [Google Scholar]
  6. Dykstra C. C., Tidwell R. R. Inhibition of topoisomerases from Pneumocystis carinii by aromatic dicationic molecules. J Protozool. 1991 Nov-Dec;38(6):78S–81S. [PubMed] [Google Scholar]
  7. Frommel T. O., Balber A. E. Flow cytofluorimetric analysis of drug accumulation by multidrug-resistant Trypanosoma brucei brucei and T. b. rhodesiense. Mol Biochem Parasitol. 1987 Nov;26(1-2):183–191. doi: 10.1016/0166-6851(87)90142-3. [DOI] [PubMed] [Google Scholar]
  8. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  9. Guide to yeast genetics and molecular biology. Methods Enzymol. 1991;194:1–863. [PubMed] [Google Scholar]
  10. Hatfield C., Kasarskis A., Staben C. Pentamidine sensitivity and resistance in Saccharomyces cerevisiae as a model for pentamidine effects on Pneumocystis carinii. J Protozool. 1991 Nov-Dec;38(6):70S–71S. [PubMed] [Google Scholar]
  11. Henikoff S., Henikoff J. G. Automated assembly of protein blocks for database searching. Nucleic Acids Res. 1991 Dec 11;19(23):6565–6572. doi: 10.1093/nar/19.23.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hughes W. T. Animal models for Pneumocystis carinii pneumonia. J Protozool. 1989 Jan-Feb;36(1):41–45. doi: 10.1111/j.1550-7408.1989.tb02688.x. [DOI] [PubMed] [Google Scholar]
  13. Hughes W. T. Prevention and treatment of Pneumocystis carinii pneumonia. Annu Rev Med. 1991;42:287–295. doi: 10.1146/annurev.me.42.020191.001443. [DOI] [PubMed] [Google Scholar]
  14. Irniger S., Braus G. H. Saturation mutagenesis of a polyadenylation signal reveals a hexanucleotide element essential for mRNA 3' end formation in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):257–261. doi: 10.1073/pnas.91.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ludewig G., Williams J. M., Li Y., Staben C. Effects of pentamidine isethionate on Saccharomyces cerevisiae. Antimicrob Agents Chemother. 1994 May;38(5):1123–1128. doi: 10.1128/aac.38.5.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nagiec M. M., Wells G. B., Lester R. L., Dickson R. C. A suppressor gene that enables Saccharomyces cerevisiae to grow without making sphingolipids encodes a protein that resembles an Escherichia coli fatty acyltransferase. J Biol Chem. 1993 Oct 15;268(29):22156–22163. [PubMed] [Google Scholar]
  17. Olson M. V., Dutchik J. E., Graham M. Y., Brodeur G. M., Helms C., Frank M., MacCollin M., Scheinman R., Frank T. Random-clone strategy for genomic restriction mapping in yeast. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7826–7830. doi: 10.1073/pnas.83.20.7826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rine J. Gene overexpression in studies of Saccharomyces cerevisiae. Methods Enzymol. 1991;194:239–251. doi: 10.1016/0076-6879(91)94019-9. [DOI] [PubMed] [Google Scholar]
  19. Salamone F. R., Cunha B. A. Update on pentamidine for the treatment of Pneumocystis carinii pneumonia. Clin Pharm. 1988 Jul;7(7):501–510. [PubMed] [Google Scholar]
  20. Schiestl R. H., Gietz R. D. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet. 1989 Dec;16(5-6):339–346. doi: 10.1007/BF00340712. [DOI] [PubMed] [Google Scholar]
  21. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sloand E., Laughon B., Armstrong M., Bartlett M. S., Blumenfeld W., Cushion M., Kalica A., Kovacs J. A., Martin W., Pitt E. The challenge of Pneumocystis carinii culture. J Eukaryot Microbiol. 1993 Mar-Apr;40(2):188–195. doi: 10.1111/j.1550-7408.1993.tb04902.x. [DOI] [PubMed] [Google Scholar]
  23. Staben C., Rabinowitz J. C. Nucleotide sequence of the Saccharomyces cerevisiae ADE3 gene encoding C1-tetrahydrofolate synthase. J Biol Chem. 1986 Apr 5;261(10):4629–4637. [PubMed] [Google Scholar]
  24. Stringer J. R., Edman J. C., Cushion M. T., Richards F. F., Watanabe J. The fungal nature of Pneumocystis. J Med Vet Mycol. 1992;30 (Suppl 1):271–278. doi: 10.1080/02681219280000961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Van Voorhis W. C. Therapy and prophylaxis of systemic protozoan infections. Drugs. 1990 Aug;40(2):176–202. doi: 10.2165/00003495-199040020-00002. [DOI] [PubMed] [Google Scholar]
  26. Walzer P. D. Pneumocystis carinii--new clinical spectrum? N Engl J Med. 1991 Jan 24;324(4):263–265. doi: 10.1056/NEJM199101243240411. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES