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. 1994 Dec 20;91(26):12907–12911. doi: 10.1073/pnas.91.26.12907

The Saccharomyces cerevisiae FKS1 (ETG1) gene encodes an integral membrane protein which is a subunit of 1,3-beta-D-glucan synthase.

C M Douglas 1, F Foor 1, J A Marrinan 1, N Morin 1, J B Nielsen 1, A M Dahl 1, P Mazur 1, W Baginsky 1, W Li 1, M el-Sherbeini 1, et al.
PMCID: PMC45549  PMID: 7528927

Abstract

In Saccharomyces cerevisiae, mutations in FKS1 confer hypersensitivity to the immunosuppressants FK506 and cyclosporin A, while mutations in ETG1 confer resistance to the cell-wall-active echinocandins (inhibitors of 1,3-beta-D-glucan synthase) and, in some cases, concomitant hypersensitivity to the chitin synthase inhibitor nikkomycin Z. The FKS1 and ETG1 genes were cloned by complementation of these phenotypes and were found to be identical. Disruption of the gene results in (i) a pronounced slow-growth phenotype, (ii) hypersensitivity to FK506 and cyclosporin A, (iii) a slight increase in sensitivity to echinocandin, and (iv) a significant reduction in 1,3-beta-D-glucan synthase activity in vitro. The nucleotide sequence encodes a 215-kDa polypeptide predicted to be an integral membrane protein with 16 transmembrane helices, consistent with previous observations that the etg1-1 mutation results in echinocandin-resistant glucan synthase activity associated with the nonextractable membrane fraction of the enzyme. These results suggest that FKS1 encodes a subunit of 1,3-beta-D-glucan synthase. The residual activity present in the disruption mutant, the nonessential nature of the gene, and results of Southern blot hybridization analysis point to the existence of a glucan synthase isozyme.

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Selected References

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

  1. Bissinger P. H., Kuchler K. Molecular cloning and expression of the Saccharomyces cerevisiae STS1 gene product. A yeast ABC transporter conferring mycotoxin resistance. J Biol Chem. 1994 Feb 11;269(6):4180–4186. [PubMed] [Google Scholar]
  2. 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]
  3. Brizuela L., Chrebet G., Bostian K. A., Parent S. A. Antifungal properties of the immunosuppressant FK-506: identification of an FK-506-responsive yeast gene distinct from FKB1. Mol Cell Biol. 1991 Sep;11(9):4616–4626. doi: 10.1128/mcb.11.9.4616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Cangelosi G. A., Martinetti G., Leigh J. A., Lee C. C., Thienes C., Theines C., Nester E. W. Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan. J Bacteriol. 1989 Mar;171(3):1609–1615. doi: 10.1128/jb.171.3.1609-1615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Douglas C. M., Marrinan J. A., Li W., Kurtz M. B. A Saccharomyces cerevisiae mutant with echinocandin-resistant 1,3-beta-D-glucan synthase. J Bacteriol. 1994 Sep;176(18):5686–5696. doi: 10.1128/jb.176.18.5686-5696.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Foor F., Parent S. A., Morin N., Dahl A. M., Ramadan N., Chrebet G., Bostian K. A., Nielsen J. B. Calcineurin mediates inhibition by FK506 and cyclosporin of recovery from alpha-factor arrest in yeast. Nature. 1992 Dec 17;360(6405):682–684. doi: 10.1038/360682a0. [DOI] [PubMed] [Google Scholar]
  9. Gewain K. M., Occi J. L., Foor F., MacNeil D. J. Vectors for generating nested deletions and facilitating subcloning G+C-rich DNA between Escherichia coli and Streptomyces sp. Gene. 1992 Sep 21;119(1):149–150. doi: 10.1016/0378-1119(92)90083-2. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hartmann E., Rapoport T. A., Lodish H. F. Predicting the orientation of eukaryotic membrane-spanning proteins. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5786–5790. doi: 10.1073/pnas.86.15.5786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Holm C., Meeks-Wagner D. W., Fangman W. L., Botstein D. A rapid, efficient method for isolating DNA from yeast. Gene. 1986;42(2):169–173. doi: 10.1016/0378-1119(86)90293-3. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Parent S. A., Nielsen J. B., Morin N., Chrebet G., Ramadan N., Dahl A. M., Hsu M. J., Bostian K. A., Foor F. Calcineurin-dependent growth of an FK506- and CsA-hypersensitive mutant of Saccharomyces cerevisiae. J Gen Microbiol. 1993 Dec;139(12):2973–2984. doi: 10.1099/00221287-139-12-2973. [DOI] [PubMed] [Google Scholar]
  15. Ram A. F., Wolters A., Ten Hoopen R., Klis F. M. A new approach for isolating cell wall mutants in Saccharomyces cerevisiae by screening for hypersensitivity to calcofluor white. Yeast. 1994 Aug;10(8):1019–1030. doi: 10.1002/yea.320100804. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Roemer T., Delaney S., Bussey H. SKN1 and KRE6 define a pair of functional homologs encoding putative membrane proteins involved in beta-glucan synthesis. Mol Cell Biol. 1993 Jul;13(7):4039–4048. doi: 10.1128/mcb.13.7.4039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rose M. D., Broach J. R. Cloning genes by complementation in yeast. Methods Enzymol. 1991;194:195–230. doi: 10.1016/0076-6879(91)94017-7. [DOI] [PubMed] [Google Scholar]
  19. Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
  20. Saier M. H., Jr Computer-aided analyses of transport protein sequences: gleaning evidence concerning function, structure, biogenesis, and evolution. Microbiol Rev. 1994 Mar;58(1):71–93. doi: 10.1128/mr.58.1.71-93.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sietsma J. H., Wessels J. G. Solubility of (1 leads to 3)-beta-D/(1 leads to 6)-beta-D-glucan in fungal walls: importance of presumed linkage between glucan and chitin. J Gen Microbiol. 1981 Jul;125(1):209–212. doi: 10.1099/00221287-125-1-209. [DOI] [PubMed] [Google Scholar]
  22. Sigal N. H., Dumont F. J. Cyclosporin A, FK-506, and rapamycin: pharmacologic probes of lymphocyte signal transduction. Annu Rev Immunol. 1992;10:519–560. doi: 10.1146/annurev.iy.10.040192.002511. [DOI] [PubMed] [Google Scholar]
  23. Sipos L., von Heijne G. Predicting the topology of eukaryotic membrane proteins. Eur J Biochem. 1993 May 1;213(3):1333–1340. doi: 10.1111/j.1432-1033.1993.tb17885.x. [DOI] [PubMed] [Google Scholar]
  24. Stanfield S. W., Ielpi L., O'Brochta D., Helinski D. R., Ditta G. S. The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB. J Bacteriol. 1988 Aug;170(8):3523–3530. doi: 10.1128/jb.170.8.3523-3530.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tang C. M., Cohen J., Holden D. W. An Aspergillus fumigatus alkaline protease mutant constructed by gene disruption is deficient in extracellular elastase activity. Mol Microbiol. 1992 Jun;6(12):1663–1671. doi: 10.1111/j.1365-2958.1992.tb00891.x. [DOI] [PubMed] [Google Scholar]
  26. Varma A., Kwon-Chung K. J. Rapid method to extract DNA from Cryptococcus neoformans. J Clin Microbiol. 1991 Apr;29(4):810–812. doi: 10.1128/jcm.29.4.810-812.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]

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