Abstract
In Saccharomyces cerevisiae, at least 12 genes are important for cells to propagate in medium containing elevated concentrations of copper salts (J. Welch, S. Fogel, C. Buchman, and M. Karin, EMBO J. 8:255-260, 1989). Complementation studies were carried out on a copper-sensitive mutation (cup14) from this group. A new yeast gene, designated SLF1, was identified as a multicopy suppressor of the cup14 mutation. Slf1 is important for the physiological process of copper sulfide (CuS) mineralization on the surface of cells cultured in medium containing copper salts. CuS mineralization causes the cells to turn brown. Disruption of SLF1, which is located close to the telomere region of chromosome IV, leads to limited copper sensitivity, and the resulting cells lack the normal brownish coloration when grown in CuSO4-containing medium. Overproduction of Slf1 in wild-type cells confers superresistance to CuSO4 and enhances the coloration of cells cultured in the presence of CuSO4. Upon addition of KCN to Cu-grown cells, the brownish coloration was bleached instantly, and copper ions were solubilized. These data are consistent with Slf1-dependent accumulation of CuS complexes on the cell surface. Disruption of SFL1 also results in loss of the ability of yeast cells to deplete Cu but not Cd ions from the growth medium, whereas overexpression enhances Ca depletion ability and the resulting deposition of CuS particles. It is proposed that Slfl participates in a copper homeostasis pathway, distinct from the Cup1 detoxification system, that leads to sulfide generation and CuS biomineralization on the cell surface. This process may coordinate with the Cup1 pathway at different copper concentrations to prevent copper-induced toxicity.
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- Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbas J., Santhanagopalan V., Blaszczynski M., Ellis W. R., Jr, Winge D. R. Conversion in the peptides coating cadmium:sulfide crystallites in Candida glabrata. J Inorg Biochem. 1992 Nov 1;48(2):95–105. doi: 10.1016/0162-0134(92)80019-r. [DOI] [PubMed] [Google Scholar]
- Butt T. R., Sternberg E. J., Gorman J. A., Clark P., Hamer D., Rosenberg M., Crooke S. T. Copper metallothionein of yeast, structure of the gene, and regulation of expression. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3332–3336. doi: 10.1073/pnas.81.11.3332. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dameron C. T., Winge D. R., George G. N., Sansone M., Hu S., Hamer D. A copper-thiolate polynuclear cluster in the ACE1 transcription factor. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6127–6131. doi: 10.1073/pnas.88.14.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dancis A., Haile D., Yuan D. S., Klausner R. D. The Saccharomyces cerevisiae copper transport protein (Ctr1p). Biochemical characterization, regulation by copper, and physiologic role in copper uptake. J Biol Chem. 1994 Oct 14;269(41):25660–25667. [PubMed] [Google Scholar]
- Dancis A., Yuan D. S., Haile D., Askwith C., Eide D., Moehle C., Kaplan J., Klausner R. D. Molecular characterization of a copper transport protein in S. cerevisiae: an unexpected role for copper in iron transport. Cell. 1994 Jan 28;76(2):393–402. doi: 10.1016/0092-8674(94)90345-x. [DOI] [PubMed] [Google Scholar]
- Eide D. J., Bridgham J. T., Zhao Z., Mattoon J. R. The vacuolar H(+)-ATPase of Saccharomyces cerevisiae is required for efficient copper detoxification, mitochondrial function, and iron metabolism. Mol Gen Genet. 1993 Nov;241(3-4):447–456. doi: 10.1007/BF00284699. [DOI] [PubMed] [Google Scholar]
- Erardi F. X., Failla M. L., Falkinham J. O., 3rd Plasmid-encoded copper resistance and precipitation by Mycobacterium scrofulaceum. Appl Environ Microbiol. 1987 Aug;53(8):1951–1954. doi: 10.1128/aem.53.8.1951-1954.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fogel S., Welch J. W. Tandem gene amplification mediates copper resistance in yeast. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5342–5346. doi: 10.1073/pnas.79.17.5342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fürst P., Hu S., Hackett R., Hamer D. Copper activates metallothionein gene transcription by altering the conformation of a specific DNA binding protein. Cell. 1988 Nov 18;55(4):705–717. doi: 10.1016/0092-8674(88)90229-2. [DOI] [PubMed] [Google Scholar]
- Goldfischer S., Schiller B., Sternlieb I. Copper in hepatocyte lysosomes of the toad, Bufo marinus L. Nature. 1970 Oct 10;228(5267):172–173. doi: 10.1038/228172a0. [DOI] [PubMed] [Google Scholar]
- Gorman J. A., Clark P. E., Lee M. C., Debouck C., Rosenberg M. Regulation of the yeast metallothionein gene. Gene. 1986;48(1):13–22. doi: 10.1016/0378-1119(86)90347-1. [DOI] [PubMed] [Google Scholar]
- Hamer D. H., Thiele D. J., Lemontt J. E. Function and autoregulation of yeast copperthionein. Science. 1985 May 10;228(4700):685–690. doi: 10.1126/science.3887570. [DOI] [PubMed] [Google Scholar]
- Hamilton R., Watanabe C. K., de Boer H. A. Compilation and comparison of the sequence context around the AUG startcodons in Saccharomyces cerevisiae mRNAs. Nucleic Acids Res. 1987 Apr 24;15(8):3581–3593. doi: 10.1093/nar/15.8.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmes J. D., Smith P. R., Evans-Gowing R., Richardson D. J., Russell D. A., Sodeau J. R. Energy-dispersive X-ray analysis of the extracellular cadmium sulfide crystallites of Klebsiella aerogenes. Arch Microbiol. 1995 Feb;163(2):143–147. doi: 10.1007/BF00381789. [DOI] [PubMed] [Google Scholar]
- Hu S., Fürst P., Hamer D. The DNA and Cu binding functions of ACE1 are interdigitated within a single domain. New Biol. 1990 Jun;2(6):544–555. [PubMed] [Google Scholar]
- Juang R. H., McCue K. F., Ow D. W. Two purine biosynthetic enzymes that are required for cadmium tolerance in Schizosaccharomyces pombe utilize cysteine sulfinate in vitro. Arch Biochem Biophys. 1993 Aug 1;304(2):392–401. doi: 10.1006/abbi.1993.1367. [DOI] [PubMed] [Google Scholar]
- Jungmann J., Reins H. A., Lee J., Romeo A., Hassett R., Kosman D., Jentsch S. MAC1, a nuclear regulatory protein related to Cu-dependent transcription factors is involved in Cu/Fe utilization and stress resistance in yeast. EMBO J. 1993 Dec 15;12(13):5051–5056. doi: 10.1002/j.1460-2075.1993.tb06198.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karin M., Najarian R., Haslinger A., Valenzuela P., Welch J., Fogel S. Primary structure and transcription of an amplified genetic locus: the CUP1 locus of yeast. Proc Natl Acad Sci U S A. 1984 Jan;81(2):337–341. doi: 10.1073/pnas.81.2.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knight S. A., Tamai K. T., Kosman D. J., Thiele D. J. Identification and analysis of a Saccharomyces cerevisiae copper homeostasis gene encoding a homeodomain protein. Mol Cell Biol. 1994 Dec;14(12):7792–7804. doi: 10.1128/mcb.14.12.7792. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin C. M., Crawford B. F., Kosman D. J. Distribution of 64Cu in Saccharomyces cerevisiae: kinetic analyses of partitioning. J Gen Microbiol. 1993 Jul;139(7):1617–1626. doi: 10.1099/00221287-139-7-1617. [DOI] [PubMed] [Google Scholar]
- Liu X. F., Culotta V. C. The requirement for yeast superoxide dismutase is bypassed through mutations in BSD2, a novel metal homeostasis gene. Mol Cell Biol. 1994 Nov;14(11):7037–7045. doi: 10.1128/mcb.14.11.7037. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miles J., Formosa T. Evidence that POB1, a Saccharomyces cerevisiae protein that binds to DNA polymerase alpha, acts in DNA metabolism in vivo. Mol Cell Biol. 1992 Dec;12(12):5724–5735. doi: 10.1128/mcb.12.12.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Minney S. F., Quirk A. V. Growth and adaptation of Saccharomyces cerevisiae at different cadmium concentrations. Microbios. 1985;42(167):37–44. [PubMed] [Google Scholar]
- Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
- Molnar J. J. Copper storage in the liver of the wild mute swan (Cygnus olor). Its possible relation to pollution of harbor waters by antifouling paints. Arch Pathol Lab Med. 1983 Dec;107(12):629–632. [PubMed] [Google Scholar]
- Nelson H., Nelson N. Disruption of genes encoding subunits of yeast vacuolar H(+)-ATPase causes conditional lethality. Proc Natl Acad Sci U S A. 1990 May;87(9):3503–3507. doi: 10.1073/pnas.87.9.3503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Philippsen P., Stotz A., Scherf C. DNA of Saccharomyces cerevisiae. Methods Enzymol. 1991;194:169–182. doi: 10.1016/0076-6879(91)94014-4. [DOI] [PubMed] [Google Scholar]
- Pringle J. R., Adams A. E., Drubin D. G., Haarer B. K. Immunofluorescence methods for yeast. Methods Enzymol. 1991;194:565–602. doi: 10.1016/0076-6879(91)94043-c. [DOI] [PubMed] [Google Scholar]
- Roberts C. J., Raymond C. K., Yamashiro C. T., Stevens T. H. Methods for studying the yeast vacuole. Methods Enzymol. 1991;194:644–661. doi: 10.1016/0076-6879(91)94047-g. [DOI] [PubMed] [Google Scholar]
- Schmitt M. E., Brown T. A., Trumpower B. L. A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res. 1990 May 25;18(10):3091–3092. doi: 10.1093/nar/18.10.3091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sikorski R. S., Boeke J. D. In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant yeast. Methods Enzymol. 1991;194:302–318. doi: 10.1016/0076-6879(91)94023-6. [DOI] [PubMed] [Google Scholar]
- Sternlieb I. Copper and the liver. Gastroenterology. 1980 Jun;78(6):1615–1628. [PubMed] [Google Scholar]
- Thiele D. J. ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene. Mol Cell Biol. 1988 Jul;8(7):2745–2752. doi: 10.1128/mcb.8.7.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thiele D. J., Hamer D. H. Tandemly duplicated upstream control sequences mediate copper-induced transcription of the Saccharomyces cerevisiae copper-metallothionein gene. Mol Cell Biol. 1986 Apr;6(4):1158–1163. doi: 10.1128/mcb.6.4.1158. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vernet T., Dignard D., Thomas D. Y. A family of yeast expression vectors containing the phage f1 intergenic region. Gene. 1987;52(2-3):225–233. doi: 10.1016/0378-1119(87)90049-7. [DOI] [PubMed] [Google Scholar]
- Welch J. W., Fogel S., Cathala G., Karin M. Industrial yeasts display tandem gene iteration at the CUP1 region. Mol Cell Biol. 1983 Aug;3(8):1353–1361. doi: 10.1128/mcb.3.8.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welch J., Fogel S., Buchman C., Karin M. The CUP2 gene product regulates the expression of the CUP1 gene, coding for yeast metallothionein. EMBO J. 1989 Jan;8(1):255–260. doi: 10.1002/j.1460-2075.1989.tb03371.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson I. A., Niman H. L., Houghten R. A., Cherenson A. R., Connolly M. L., Lerner R. A. The structure of an antigenic determinant in a protein. Cell. 1984 Jul;37(3):767–778. doi: 10.1016/0092-8674(84)90412-4. [DOI] [PubMed] [Google Scholar]