Abstract
A novel, potent, semisynthetic pneumocandin, L-733,560, was used to isolate a resistant mutant in Saccharomyces cerevisiae. This compound, like other pneumocandins and echinocandins, inhibits 1,3-beta-D-glucan synthase from Candida albicans (F.A. Bouffard, R.A. Zambias, J. F. Dropinski, J.M. Balkovec, M.L. Hammond, G.K. Abruzzo, K.F. Bartizal, J.A. Marrinan, M. B. Kurtz, D.C. McFadden, K.H. Nollstadt, M.A. Powles, and D.M. Schmatz, J. Med. Chem. 37:222-225, 1994). Glucan synthesis catalyzed by a crude membrane fraction prepared from the S. cerevisiae mutant R560-1C was resistant to inhibition by L-733,560. The nearly 50-fold increase in the 50% inhibitory concentration against glucan synthase was commensurate with the increase in whole-cell resistance. R560-1C was cross-resistant to other inhibitors of C. albicans 1,3-beta-D-glucan synthase (aculeacin A, dihydropapulacandin, and others) but not to compounds with different modes of action. Genetic analysis revealed that enzyme and whole-cell pneumocandin resistance was due to a single mutant gene, designated etg1-1 (echinocandin target gene 1), which was semidominant in heterozygous diploids. The etg1-1 mutation did not confer enhanced ability to metabolize L-733,560 and had no effect on the membrane-bound enzymes chitin synthase I and squalene synthase. Alkali-soluble beta-glucan synthesized by crude microsomes from R560-1C was indistinguishable from the wild-type product. 1,3-beta-D-Glucan synthase activity from R560-1C was fractionated with NaCl and Tergitol NP-40; reconstitution with fractions from wild-type membranes revealed that drug resistance is associated with the insoluble membrane fraction. We propose that the etg1-1 mutant gene encodes a subunit of the 1,3-beta-D-glucan synthase complex.
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Selected References
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- Alberts A. W., Chen J., Kuron G., Hunt V., Huff J., Hoffman C., Rothrock J., Lopez M., Joshua H., Harris E. Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-coenzyme A reductase and a cholesterol-lowering agent. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3957–3961. doi: 10.1073/pnas.77.7.3957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Balzi E., Chen W., Ulaszewski S., Capieaux E., Goffeau A. The multidrug resistance gene PDR1 from Saccharomyces cerevisiae. J Biol Chem. 1987 Dec 15;262(35):16871–16879. [PubMed] [Google Scholar]
- Bartizal K., Abruzzo G., Trainor C., Krupa D., Nollstadt K., Schmatz D., Schwartz R., Hammond M., Balkovec J., Vanmiddlesworth F. In vitro antifungal activities and in vivo efficacies of 1,3-beta-D-glucan synthesis inhibitors L-671,329, L-646,991, tetrahydroechinocandin B, and L-687,781, a papulacandin. Antimicrob Agents Chemother. 1992 Aug;36(8):1648–1657. doi: 10.1128/aac.36.8.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beaulieu D., Tang J., Zeckner D. J., Parr T. R., Jr Correlation of cilofungin in vivo efficacy with its activity against Aspergillus fumigatus (1,3)-beta-D-glucan synthase. FEMS Microbiol Lett. 1993 Apr 1;108(2):133–137. doi: 10.1111/j.1574-6968.1993.tb06088.x. [DOI] [PubMed] [Google Scholar]
- Beauvais A., Drake R., Ng K., Diaquin M., Latgé J. P. Characterization of the 1,3-beta-glucan synthase of Aspergillus fumigatus. J Gen Microbiol. 1993 Dec;139(12):3071–3078. doi: 10.1099/00221287-139-12-3071. [DOI] [PubMed] [Google Scholar]
- Bensadoun A., Weinstein D. Assay of proteins in the presence of interfering materials. Anal Biochem. 1976 Jan;70(1):241–250. doi: 10.1016/s0003-2697(76)80064-4. [DOI] [PubMed] [Google Scholar]
- Bergstrom J. D., Kurtz M. M., Rew D. J., Amend A. M., Karkas J. D., Bostedor R. G., Bansal V. S., Dufresne C., VanMiddlesworth F. L., Hensens O. D. Zaragozic acids: a family of fungal metabolites that are picomolar competitive inhibitors of squalene synthase. Proc Natl Acad Sci U S A. 1993 Jan 1;90(1):80–84. doi: 10.1073/pnas.90.1.80. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
- Boone C., Sommer S. S., Hensel A., Bussey H. Yeast KRE genes provide evidence for a pathway of cell wall beta-glucan assembly. J Cell Biol. 1990 May;110(5):1833–1843. doi: 10.1083/jcb.110.5.1833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borgia P. T., Dodge C. L. Characterization of Aspergillus nidulans mutants deficient in cell wall chitin or glucan. J Bacteriol. 1992 Jan;174(2):377–383. doi: 10.1128/jb.174.2.377-383.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bouffard F. A., Zambias R. A., Dropinski J. F., Balkovec J. M., Hammond M. L., Abruzzo G. K., Bartizal K. F., Marrinan J. A., Kurtz M. B., McFadden D. C. Synthesis and antifungal activity of novel cationic pneumocandin B(o) derivatives. J Med Chem. 1994 Jan 21;37(2):222–225. doi: 10.1021/jm00028a003. [DOI] [PubMed] [Google Scholar]
- Bulawa C. E. Genetics and molecular biology of chitin synthesis in fungi. Annu Rev Microbiol. 1993;47:505–534. doi: 10.1146/annurev.mi.47.100193.002445. [DOI] [PubMed] [Google Scholar]
- Bussey H., Saville D., Hutchins K., Palfree R. G. Binding of yeast killer toxin to a cell wall receptor on sensitive Saccharomyces cerevisiae. J Bacteriol. 1979 Dec;140(3):888–892. doi: 10.1128/jb.140.3.888-892.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cabib E., Bowers B., Sburlati A., Silverman S. J. Fungal cell wall synthesis: the construction of a biological structure. Microbiol Sci. 1988 Dec;5(12):370–375. [PubMed] [Google Scholar]
- Cabib E., Kang M. S., Au-Young J. Chitin synthase from Saccharomyces cerevisiae. Methods Enzymol. 1987;138:643–649. doi: 10.1016/0076-6879(87)38058-9. [DOI] [PubMed] [Google Scholar]
- Cabib E., Kang M. S. Fungal 1,3-beta-glucan synthase. Methods Enzymol. 1987;138:637–642. doi: 10.1016/0076-6879(87)38057-7. [DOI] [PubMed] [Google Scholar]
- Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
- Cassone A., Mason R. E., Kerridge D. Lysis of growing yeast-form cells of Candida albicans by echinocandin: a cytological study. Sabouraudia. 1981 Jun;19(2):97–110. [PubMed] [Google Scholar]
- Font de Mora J., Gil R., Sentandreu R., Herrero E. Isolation and characterization of Saccharomyces cerevisiae mutants resistant to aculeacin A. Antimicrob Agents Chemother. 1991 Dec;35(12):2596–2601. doi: 10.1128/aac.35.12.2596. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong Z., Mann P., Brown N. H., Tran L. E., Shaw K. J., Hare R. S., DiDomenico B. Cloning and characterization of KNR4, a yeast gene involved in (1,3)-beta-glucan synthesis. Mol Cell Biol. 1994 Feb;14(2):1017–1025. doi: 10.1128/mcb.14.2.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horn W. S., Smith J. L., Bills G. F., Raghoobar S. L., Helms G. L., Kurtz M. B., Marrinan J. A., Frommer B. R., Thornton R. A., Mandala S. M. Sphingofungins E and F: novel serinepalmitoyl transferase inhibitors from Paecilomyces variotii. J Antibiot (Tokyo) 1992 Oct;45(10):1692–1696. doi: 10.7164/antibiotics.45.1692. [DOI] [PubMed] [Google Scholar]
- Hutchins K., Bussey H. Cell wall receptor for yeast killer toxin: involvement of (1 leads to 6)-beta-D-glucan. J Bacteriol. 1983 Apr;154(1):161–169. doi: 10.1128/jb.154.1.161-169.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kang M. S., Cabib E. Regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (1----3)-beta-D-glucan synthase. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5808–5812. doi: 10.1073/pnas.83.16.5808. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kasahara S., Yamada H., Mio T., Shiratori Y., Miyamoto C., Yabe T., Nakajima T., Ichishima E., Furuichi Y. Cloning of the Saccharomyces cerevisiae gene whose overexpression overcomes the effects of HM-1 killer toxin, which inhibits beta-glucan synthesis. J Bacteriol. 1994 Mar;176(5):1488–1499. doi: 10.1128/jb.176.5.1488-1499.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller-Schierlein W., Widmer J. Stoffwechselprodukte von Mikroorganismen Uber die aromatische Aminosäure des Echinocandins B: 3,4-Dihydroxyhomotyrosin. Helv Chim Acta. 1976 Sep 29;59(6):2021–2031. doi: 10.1002/hlca.19760590615. [DOI] [PubMed] [Google Scholar]
- Kurtz M. B., Heath I. B., Marrinan J., Dreikorn S., Onishi J., Douglas C. Morphological effects of lipopeptides against Aspergillus fumigatus correlate with activities against (1,3)-beta-D-glucan synthase. Antimicrob Agents Chemother. 1994 Jul;38(7):1480–1489. doi: 10.1128/aac.38.7.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meaden P., Hill K., Wagner J., Slipetz D., Sommer S. S., Bussey H. The yeast KRE5 gene encodes a probable endoplasmic reticulum protein required for (1----6)-beta-D-glucan synthesis and normal cell growth. Mol Cell Biol. 1990 Jun;10(6):3013–3019. doi: 10.1128/mcb.10.6.3013. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mizuno K., Yagi A., Satoi S., Takada M., Hayashi M. Studies on aculeacin. I. Isolation and characterization of aculeacin A. J Antibiot (Tokyo) 1977 Apr;30(4):297–302. doi: 10.7164/antibiotics.30.297. [DOI] [PubMed] [Google Scholar]
- Odds F. C., Cheesman S. L., Abbott A. B. Antifungal effects of fluconazole (UK 49858), a new triazole antifungal, in vitro. J Antimicrob Chemother. 1986 Oct;18(4):473–478. doi: 10.1093/jac/18.4.473. [DOI] [PubMed] [Google Scholar]
- Orlean P. A. (1,3)-beta-D-Glucan synthase from budding and filamentous cultures of the dimorphic fungus Candida albicans. Eur J Biochem. 1982 Oct;127(2):397–403. doi: 10.1111/j.1432-1033.1982.tb06885.x. [DOI] [PubMed] [Google Scholar]
- Petranyi G., Ryder N. S., Stütz A. Allylamine derivatives: new class of synthetic antifungal agents inhibiting fungal squalene epoxidase. Science. 1984 Jun 15;224(4654):1239–1241. doi: 10.1126/science.6547247. [DOI] [PubMed] [Google Scholar]
- Ribas J. C., Diaz M., Duran A., Perez P. Isolation and characterization of Schizosaccharomyces pombe mutants defective in cell wall (1-3)beta-D-glucan. J Bacteriol. 1991 Jun;173(11):3456–3462. doi: 10.1128/jb.173.11.3456-3462.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roemer T., Bussey H. Yeast beta-glucan synthesis: KRE6 encodes a predicted type II membrane protein required for glucan synthesis in vivo and for glucan synthase activity in vitro. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11295–11299. doi: 10.1073/pnas.88.24.11295. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roncero C., Valdivieso M. H., Ribas J. C., Durán A. Effect of calcofluor white on chitin synthases from Saccharomyces cerevisiae. J Bacteriol. 1988 Apr;170(4):1945–1949. doi: 10.1128/jb.170.4.1945-1949.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roncero C., Valdivieso M. H., Ribas J. C., Durán A. Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white. J Bacteriol. 1988 Apr;170(4):1950–1954. doi: 10.1128/jb.170.4.1950-1954.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sawistowska-Schröder E. T., Kerridge D., Perry H. Echinocandin inhibition of 1,3-beta-D-glucan synthase from Candida albicans. FEBS Lett. 1984 Jul 23;173(1):134–138. doi: 10.1016/0014-5793(84)81032-7. [DOI] [PubMed] [Google Scholar]
- Schmatz D. M., Romancheck M. A., Pittarelli L. A., Schwartz R. E., Fromtling R. A., Nollstadt K. H., Vanmiddlesworth F. L., Wilson K. E., Turner M. J. Treatment of Pneumocystis carinii pneumonia with 1,3-beta-glucan synthesis inhibitors. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5950–5954. doi: 10.1073/pnas.87.15.5950. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz R. E., Sesin D. F., Joshua H., Wilson K. E., Kempf A. J., Goklen K. A., Kuehner D., Gailliot P., Gleason C., White R. Pneumocandins from Zalerion arboricola. I. Discovery and isolation. J Antibiot (Tokyo) 1992 Dec;45(12):1853–1866. doi: 10.7164/antibiotics.45.1853. [DOI] [PubMed] [Google Scholar]
- Selitrennikoff C. P. Use of a temperature-sensitive, protoplast-forming Neurospora crassa strain for the detection of antifungal antibiotics. Antimicrob Agents Chemother. 1983 May;23(5):757–765. doi: 10.1128/aac.23.5.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shematek E. M., Braatz J. A., Cabib E. Biosynthesis of the yeast cell wall. I. Preparation and properties of beta-(1 leads to 3)glucan synthetase. J Biol Chem. 1980 Feb 10;255(3):888–894. [PubMed] [Google Scholar]
- Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
- Tang J., Parr T. R., Jr W-1 solubilization and kinetics of inhibition by cilofungin of Candida albicans (1,3)-beta-D-glucan synthase. Antimicrob Agents Chemother. 1991 Jan;35(1):99–103. doi: 10.1128/aac.35.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
- Valdivieso M. H., Mol P. C., Shaw J. A., Cabib E., Durán A. CAL1, a gene required for activity of chitin synthase 3 in Saccharomyces cerevisiae. J Cell Biol. 1991 Jul;114(1):101–109. doi: 10.1083/jcb.114.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VanMiddlesworth F., Omstead M. N., Schmatz D., Bartizal K., Fromtling R., Bills G., Nollstadt K., Honeycutt S., Zweerink M., Garrity G. L-687,781, a new member of the papulacandin family of beta-1,3-D-glucan synthesis inhibitors. I. Fermentation, isolation, and biological activity. J Antibiot (Tokyo) 1991 Jan;44(1):45–51. doi: 10.7164/antibiotics.44.45. [DOI] [PubMed] [Google Scholar]
- Yamaguchi H., Hiratani T., Iwata K., Yamamoto Y. Studies on the mechanism of antifungal action of aculeacin A. J Antibiot (Tokyo) 1982 Feb;35(2):210–219. doi: 10.7164/antibiotics.35.210. [DOI] [PubMed] [Google Scholar]