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. 1994 Oct;14(10):6597–6606. doi: 10.1128/mcb.14.10.6597

Two FK506 resistance-conferring genes in Saccharomyces cerevisiae, TAT1 and TAT2, encode amino acid permeases mediating tyrosine and tryptophan uptake.

A Schmidt 1, M N Hall 1, A Koller 1
PMCID: PMC359189  PMID: 7523855

Abstract

The macrocyclic lactone FK506 exerts immunosuppressive effects on T lymphocytes by interfering with signal transduction leading to T-cell activation and also inhibits the growth of eukaryotic microorganisms, including Saccharomyces cerevisiae. We reported previously that an FK506-sensitive target in S. cerevisiae is required for amino acid import and that overexpression of two new genes, TAT1 and TAT2 (formerly called TAP1 and TAP2), confers resistance to the drug. Here we report that TAT1 and TAT2 encode novel members of the yeast amino acid permease family composed of integral membrane proteins that share 30 to 40% identity. TAT1 is the tyrosine high-affinity transporter, which also mediates low-affinity or low-capacity uptake of tryptophan. TAT2 is the tryptophan high-affinity transporter. FK506 does not reduce the levels of TAT1 and TAT2 transcripts, indicating that the inhibition of amino acid transport by the drug is posttranscriptional.

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Selected References

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  1. Ahmad M., Bussey H. Yeast arginine permease: nucleotide sequence of the CAN1 gene. Curr Genet. 1986;10(8):587–592. doi: 10.1007/BF00418125. [DOI] [PubMed] [Google Scholar]
  2. Aldrich T. L., Di Segni G., McConaughy B. L., Keen N. J., Whelen S., Hall B. D. Structure of the yeast TAP1 protein: dependence of transcription activation on the DNA context of the target gene. Mol Cell Biol. 1993 Jun;13(6):3434–3444. doi: 10.1128/mcb.13.6.3434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arndt K., Fink G. R. GCN4 protein, a positive transcription factor in yeast, binds general control promoters at all 5' TGACTC 3' sequences. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8516–8520. doi: 10.1073/pnas.83.22.8516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braus G. H. Aromatic amino acid biosynthesis in the yeast Saccharomyces cerevisiae: a model system for the regulation of a eukaryotic biosynthetic pathway. Microbiol Rev. 1991 Sep;55(3):349–370. doi: 10.1128/mr.55.3.349-370.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Courchesne W. E., Magasanik B. Ammonia regulation of amino acid permeases in Saccharomyces cerevisiae. Mol Cell Biol. 1983 Apr;3(4):672–683. doi: 10.1128/mcb.3.4.672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crabeel M., Grenson M. Regulation of histidine uptake by specific feedback inhibition of two histidine permeases in Saccharomyces cerevisiae. Eur J Biochem. 1970 May 1;14(1):197–204. doi: 10.1111/j.1432-1033.1970.tb00278.x. [DOI] [PubMed] [Google Scholar]
  7. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Domdey H., Apostol B., Lin R. J., Newman A., Brody E., Abelson J. Lariat structures are in vivo intermediates in yeast pre-mRNA splicing. Cell. 1984 Dec;39(3 Pt 2):611–621. doi: 10.1016/0092-8674(84)90468-9. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Greasham R. L., Moat A. G. Amino acid transport in a polyaromatic amino acid auxotroph of Saccharomyces cerevisiae. J Bacteriol. 1973 Sep;115(3):975–981. doi: 10.1128/jb.115.3.975-981.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grenson M. Inactivation-reactivation process and repression of permease formation regulate several ammonia-sensitive permeases in the yeast Saccharomyces cerevisiae. Eur J Biochem. 1983 Jun 1;133(1):135–139. doi: 10.1111/j.1432-1033.1983.tb07438.x. [DOI] [PubMed] [Google Scholar]
  12. Grenson M. Study of the positive control of the general amino-acid permease and other ammonia-sensitive uptake systems by the product of the NPR1 gene in the yeast Saccharomyces cerevisiae. Eur J Biochem. 1983 Jun 1;133(1):141–144. doi: 10.1111/j.1432-1033.1983.tb07439.x. [DOI] [PubMed] [Google Scholar]
  13. Heitman J., Koller A., Kunz J., Henriquez R., Schmidt A., Movva N. R., Hall M. N. The immunosuppressant FK506 inhibits amino acid import in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Aug;13(8):5010–5019. doi: 10.1128/mcb.13.8.5010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heitman J., Movva N. R., Hiestand P. C., Hall M. N. FK 506-binding protein proline rotamase is a target for the immunosuppressive agent FK 506 in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1948–1952. doi: 10.1073/pnas.88.5.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hill D. E., Hope I. A., Macke J. P., Struhl K. Saturation mutagenesis of the yeast his3 regulatory site: requirements for transcriptional induction and for binding by GCN4 activator protein. Science. 1986 Oct 24;234(4775):451–457. doi: 10.1126/science.3532321. [DOI] [PubMed] [Google Scholar]
  16. Hinnebusch A. G., Fink G. R. Positive regulation in the general amino acid control of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5374–5378. doi: 10.1073/pnas.80.17.5374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hinnebusch A. G., Fink G. R. Repeated DNA sequences upstream from HIS1 also occur at several other co-regulated genes in Saccharomyces cerevisiae. J Biol Chem. 1983 Apr 25;258(8):5238–5247. [PubMed] [Google Scholar]
  18. Hoffmann W. Molecular characterization of the CAN1 locus in Saccharomyces cerevisiae. A transmembrane protein without N-terminal hydrophobic signal sequence. J Biol Chem. 1985 Sep 25;260(21):11831–11837. [PubMed] [Google Scholar]
  19. Hope I. A., Struhl K. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast. Cell. 1985 Nov;43(1):177–188. doi: 10.1016/0092-8674(85)90022-4. [DOI] [PubMed] [Google Scholar]
  20. Horák J. Amino acid transport in eucaryotic microorganisms. Biochim Biophys Acta. 1986 Dec 22;864(3-4):223–256. doi: 10.1016/0304-4157(86)90001-8. [DOI] [PubMed] [Google Scholar]
  21. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jauniaux J. C., Grenson M. GAP1, the general amino acid permease gene of Saccharomyces cerevisiae. Nucleotide sequence, protein similarity with the other bakers yeast amino acid permeases, and nitrogen catabolite repression. Eur J Biochem. 1990 May 31;190(1):39–44. doi: 10.1111/j.1432-1033.1990.tb15542.x. [DOI] [PubMed] [Google Scholar]
  23. Jauniaux J. C., Vandenbol M., Vissers S., Broman K., Grenson M. Nitrogen catabolite regulation of proline permease in Saccharomyces cerevisiae. Cloning of the PUT4 gene and study of PUT4 RNA levels in wild-type and mutant strains. Eur J Biochem. 1987 May 4;164(3):601–606. doi: 10.1111/j.1432-1033.1987.tb11169.x. [DOI] [PubMed] [Google Scholar]
  24. Kunz J., Hall M. N. Cyclosporin A, FK506 and rapamycin: more than just immunosuppression. Trends Biochem Sci. 1993 Sep;18(9):334–338. doi: 10.1016/0968-0004(93)90069-y. [DOI] [PubMed] [Google Scholar]
  25. Larimore F. S., Roon R. J. Possible site-specific reagent for the general amino acid transport system of Saccharomyces cerevisiae. Biochemistry. 1978 Feb 7;17(3):431–436. doi: 10.1021/bi00596a008. [DOI] [PubMed] [Google Scholar]
  26. McCusker J. H., Haber J. E. Mutations in Saccharomyces cerevisiae which confer resistance to several amino acid analogs. Mol Cell Biol. 1990 Jun;10(6):2941–2949. doi: 10.1128/mcb.10.6.2941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miozzari G., Niederberger P., Hütter R. Action of tryptophan analogues in Saccharomyces cerevisiae. Arch Microbiol. 1977 Dec 15;115(3):307–316. doi: 10.1007/BF00446457. [DOI] [PubMed] [Google Scholar]
  28. Mösch H. U., Scheier B., Lahti R., Mäntsäla P., Braus G. H. Transcriptional activation of yeast nucleotide biosynthetic gene ADE4 by GCN4. J Biol Chem. 1991 Oct 25;266(30):20453–20456. [PubMed] [Google Scholar]
  29. Oliver S. G., van der Aart Q. J., Agostoni-Carbone M. L., Aigle M., Alberghina L., Alexandraki D., Antoine G., Anwar R., Ballesta J. P., Benit P. The complete DNA sequence of yeast chromosome III. Nature. 1992 May 7;357(6373):38–46. doi: 10.1038/357038a0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  33. Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. doi: 10.1016/0076-6879(91)94004-v. [DOI] [PubMed] [Google Scholar]
  34. Stotz A., Müller P. P., Linder P. Regulation of the ADE2 gene from Saccharomyces cerevisiae. Curr Genet. 1993 Dec;24(6):472–480. doi: 10.1007/BF00351708. [DOI] [PubMed] [Google Scholar]
  35. Sychrova H., Chevallier M. R. Cloning and sequencing of the Saccharomyces cerevisiae gene LYP1 coding for a lysine-specific permease. Yeast. 1993 Jul;9(7):771–782. doi: 10.1002/yea.320090711. [DOI] [PubMed] [Google Scholar]
  36. Tanaka J., Fink G. R. The histidine permease gene (HIP1) of Saccharomyces cerevisiae. Gene. 1985;38(1-3):205–214. doi: 10.1016/0378-1119(85)90219-7. [DOI] [PubMed] [Google Scholar]
  37. Vandenbol M., Jauniaux J. C., Grenson M. Nucleotide sequence of the Saccharomyces cerevisiae PUT4 proline-permease-encoding gene: similarities between CAN1, HIP1 and PUT4 permeases. Gene. 1989 Nov 15;83(1):153–159. doi: 10.1016/0378-1119(89)90413-7. [DOI] [PubMed] [Google Scholar]
  38. Vandenbol M., Jauniaux J. C., Grenson M. The Saccharomyces cerevisiae NPR1 gene required for the activity of ammonia-sensitive amino acid permeases encodes a protein kinase homologue. Mol Gen Genet. 1990 Jul;222(2-3):393–399. doi: 10.1007/BF00633845. [DOI] [PubMed] [Google Scholar]
  39. Vandenbol M., Jauniaux J. C., Vissers S., Grenson M. Isolation of the NPR1 gene responsible for the reactivation of ammonia-sensitive amino-acid permeases in Saccharomyces cerevisiae. RNA analysis and gene dosage effects. Eur J Biochem. 1987 May 4;164(3):607–612. doi: 10.1111/j.1432-1033.1987.tb11170.x. [DOI] [PubMed] [Google Scholar]
  40. Wiame J. M., Grenson M., Arst H. N., Jr Nitrogen catabolite repression in yeasts and filamentous fungi. Adv Microb Physiol. 1985;26:1–88. doi: 10.1016/s0065-2911(08)60394-x. [DOI] [PubMed] [Google Scholar]
  41. Woodward J. R., Kornberg H. L. Membrane proteins associated with amino acid transport by yeast (Saccharomyces cerevisiae). Biochem J. 1980 Nov 15;192(2):659–664. doi: 10.1042/bj1920659. [DOI] [PMC free article] [PubMed] [Google Scholar]

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