• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of geneticsGeneticsCurrent IssueInformation for AuthorsEditorial BoardSubscribeSubmit a Manuscript
Genetics. Apr 2003; 163(4): 1287–1298.
PMCID: PMC1462505

Mode of selection and experimental evolution of antifungal drug resistance in Saccharomyces cerevisiae.

Abstract

We show that mode of selection, degree of dominance of mutations, and ploidy are determining factors in the evolution of resistance to the antifungal drug fluconazole in yeast. In experiment 1, yeast populations were subjected to a stepwise increase in fluconazole concentration over 400 generations. Under this regimen, two mutations in the same two chromosomal regions rose to high frequency in parallel in three replicate populations. These mutations were semidominant and additive in their effect on resistance. The first of these mutations mapped to PDR1 and resulted in the overexpression of the ABC transporter genes PDR5 and SNQ2. These mutations had an unexpected pleiotropic effect of reducing the residual ability of the wild type to reproduce at the highest concentrations of fluconazole. In experiment 2, yeast populations were subjected to a single high concentration of fluconazole. Under this regimen, a single recessive mutation appeared in each of three replicate populations. In a genome-wide screen of approximately 4700 viable deletion strains, 13 were classified as resistant to fluconazole (ERG3, ERG6, YMR102C, YMR099C, YPL056C, ERG28, OSH1, SCS2, CKA2, SML1, YBR147W, YGR283C, and YLR407W). The mutations in experiment 2 all mapped to ERG3 and resulted in the overexpression of the gene encoding the drug target ERG11, but not PDR5 and SNQ2. Diploid hybrids from experiments 1 and 2 were less fit than the parents in the presence of fluconazole. In a variation of experiment 2, haploids showed a higher frequency of resistance than diploids, suggesting that degree of dominance and ploidy are important factors in the evolution of antifungal drug resistance.

Full Text

The Full Text of this article is available as a PDF (222K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andersson DI, Levin BR. The biological cost of antibiotic resistance. Curr Opin Microbiol. 1999 Oct;2(5):489–493. [PubMed]
  • Beh CT, Cool L, Phillips J, Rine J. Overlapping functions of the yeast oxysterol-binding protein homologues. Genetics. 2001 Mar;157(3):1117–1140. [PMC free article] [PubMed]
  • Cowen LE, Sanglard D, Calabrese D, Sirjusingh C, Anderson JB, Kohn LM. Evolution of drug resistance in experimental populations of Candida albicans. J Bacteriol. 2000 Mar;182(6):1515–1522. [PMC free article] [PubMed]
  • Cowen LE, Kohn LM, Anderson JB. Divergence in fitness and evolution of drug resistance in experimental populations of Candida albicans. J Bacteriol. 2001 May;183(10):2971–2978. [PMC free article] [PubMed]
  • Cowen Leah E, Nantel André, Whiteway Malcolm S, Thomas David Y, Tessier Daniel C, Kohn Linda M, Anderson James B. Population genomics of drug resistance in Candida albicans. Proc Natl Acad Sci U S A. 2002 Jul 9;99(14):9284–9289. [PMC free article] [PubMed]
  • Cowen Leah E, Anderson James B, Kohn Linda M. Evolution of drug resistance in Candida albicans. Annu Rev Microbiol. 2002;56:139–165. [PubMed]
  • DeRisi J, van den Hazel B, Marc P, Balzi E, Brown P, Jacq C, Goffeau A. Genome microarray analysis of transcriptional activation in multidrug resistance yeast mutants. FEBS Lett. 2000 Mar 24;470(2):156–160. [PubMed]
  • Gachotte D, Eckstein J, Barbuch R, Hughes T, Roberts C, Bard M. A novel gene conserved from yeast to humans is involved in sterol biosynthesis. J Lipid Res. 2001 Jan;42(1):150–154. [PubMed]
  • Georgopapadakou NH, Walsh TJ. Human mycoses: drugs and targets for emerging pathogens. Science. 1994 Apr 15;264(5157):371–373. [PubMed]
  • Hughes TR, Marton MJ, Jones AR, Roberts CJ, Stoughton R, Armour CD, Bennett HA, Coffey E, Dai H, He YD, et al. Functional discovery via a compendium of expression profiles. Cell. 2000 Jul 7;102(1):109–126. [PubMed]
  • Kagiwada S, Hosaka K, Murata M, Nikawa J, Takatsuki A. The Saccharomyces cerevisiae SCS2 gene product, a homolog of a synaptobrevin-associated protein, is an integral membrane protein of the endoplasmic reticulum and is required for inositol metabolism. J Bacteriol. 1998 Apr;180(7):1700–1708. [PMC free article] [PubMed]
  • Kolaczkowska Anna, Goffeau Andre. Regulation of pleiotropic drug resistance in yeast. Drug Resist Updat. 1999 Dec;2(6):403–414. [PubMed]
  • Kolaczkowska A, Kolaczkowski M, Delahodde A, Goffeau A. Functional dissection of Pdr1p, a regulator of multidrug resistance in Saccharomyces cerevisiae. Mol Genet Genomics. 2002 Mar;267(1):96–106. [PubMed]
  • Levin BR, Perrot V, Walker N. Compensatory mutations, antibiotic resistance and the population genetics of adaptive evolution in bacteria. Genetics. 2000 Mar;154(3):985–997. [PMC free article] [PubMed]
  • Lupetti Antonella, Danesi Romano, Campa Mario, Del Tacca Mario, Kelly Steven. Molecular basis of resistance to azole antifungals. Trends Mol Med. 2002 Feb;8(2):76–81. [PubMed]
  • Marchetti O, Entenza JM, Sanglard D, Bille J, Glauser MP, Moreillon P. Fluconazole plus cyclosporine: a fungicidal combination effective against experimental endocarditis due to Candida albicans. Antimicrob Agents Chemother. 2000 Nov;44(11):2932–2938. [PMC free article] [PubMed]
  • Sanglard D, Ischer F, Calabrese D, Micheli M, Bille J. Multiple resistance mechanisms to azole antifungals in yeast clinical isolates. Drug Resist Updat. 1998;1(4):255–265. [PubMed]
  • Marr KA, Rustad TR, Rex JH, White TC. The trailing end point phenotype in antifungal susceptibility testing is pH dependent. Antimicrob Agents Chemother. 1999 Jun;43(6):1383–1386. [PMC free article] [PubMed]
  • Sato M, Fujisaki S, Sato K, Nishimura Y, Nakano A. Yeast Saccharomyces cerevisiae has two cis-prenyltransferases with different properties and localizations. Implication for their distinct physiological roles in dolichol synthesis. Genes Cells. 2001 Jun;6(6):495–506. [PubMed]
  • Orr HA, Otto SP. Does diploidy increase the rate of adaptation? Genetics. 1994 Apr;136(4):1475–1480. [PMC free article] [PubMed]
  • Sherman F, Hicks J. Micromanipulation and dissection of asci. Methods Enzymol. 1991;194:21–37. [PubMed]
  • Taylor JW, Geiser DM, Burt A, Koufopanou V. The evolutionary biology and population genetics underlying fungal strain typing. Clin Microbiol Rev. 1999 Jan;12(1):126–146. [PMC free article] [PubMed]
  • Wach A, Brachat A, Pöhlmann R, Philippsen P. New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast. 1994 Dec;10(13):1793–1808. [PubMed]
  • Rex JH, Pfaller MA, Walsh TJ, Chaturvedi V, Espinel-Ingroff A, Ghannoum MA, Gosey LL, Odds FC, Rinaldi MG, Sheehan DJ, et al. Antifungal susceptibility testing: practical aspects and current challenges. Clin Microbiol Rev. 2001 Oct;14(4):643–contents. [PMC free article] [PubMed]
  • Reyes Guadalupe, Ghannoum Mahmoud A. Antifungal susceptibility testing of yeasts: uses and limitations. Drug Resist Updat. 2000 Feb;3(1):14–19. [PubMed]
  • Reynolds TB, Fink GR. Bakers' yeast, a model for fungal biofilm formation. Science. 2001 Feb 2;291(5505):878–881. [PubMed]

Articles from Genetics are provided here courtesy of Genetics Society of America

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...