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. 1998 Dec 1;17(23):6924–6931. doi: 10.1093/emboj/17.23.6924

The TOR nutrient signalling pathway phosphorylates NPR1 and inhibits turnover of the tryptophan permease.

A Schmidt 1, T Beck 1, A Koller 1, J Kunz 1, M N Hall 1
PMCID: PMC1171040  PMID: 9843498

Abstract

The Saccharomyces cerevisiae targets of rapamycin, TOR1 and TOR2, signal activation of cell growth in response to nutrient availability. Loss of TOR or rapamycin treatment causes yeast cells to arrest growth in early G1 and to express several other physiological properties of starved (G0) cells. As part of this starvation response, high affinity amino acid permeases such as the tryptophan permease TAT2 are targeted to the vacuole and degraded. Here we show that the TOR signalling pathway phosphorylates the Ser/Thr kinase NPR1 and thereby inhibits the starvation-induced turnover of TAT2. Overexpression of NPR1 inhibits growth and induces the degradation of TAT2, whereas loss of NPR1 confers resistance to rapamycin and to FK506, an inhibitor of amino acid import. NPR1 is controlled by TOR and the type 2A phosphatase-associated protein TAP42. First, overexpression of NPR1 is toxic only when TOR function is reduced. Secondly, NPR1 is rapidly dephosphorylated in the absence of TOR. Thirdly, NPR1 dephosphorylation does not occur in a rapamycin-resistant tap42 mutant. Thus, the TOR nutrient signalling pathway also controls growth by inhibiting a stationary phase (G0) programme. The control of NPR1 by TOR is analogous to the control of p70 s6 kinase and 4E-BP1 by mTOR in mammalian cells.

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Selected References

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  1. Barbet N. C., Schneider U., Helliwell S. B., Stansfield I., Tuite M. F., Hall M. N. TOR controls translation initiation and early G1 progression in yeast. Mol Biol Cell. 1996 Jan;7(1):25–42. doi: 10.1091/mbc.7.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Begum N., Ragolia L. cAMP counter-regulates insulin-mediated protein phosphatase-2A inactivation in rat skeletal muscle cells. J Biol Chem. 1996 Dec 6;271(49):31166–31171. doi: 10.1074/jbc.271.49.31166. [DOI] [PubMed] [Google Scholar]
  3. Beretta L., Gingras A. C., Svitkin Y. V., Hall M. N., Sonenberg N. Rapamycin blocks the phosphorylation of 4E-BP1 and inhibits cap-dependent initiation of translation. EMBO J. 1996 Feb 1;15(3):658–664. [PMC free article] [PubMed] [Google Scholar]
  4. Bickle M., Delley P. A., Schmidt A., Hall M. N. Cell wall integrity modulates RHO1 activity via the exchange factor ROM2. EMBO J. 1998 Apr 15;17(8):2235–2245. doi: 10.1093/emboj/17.8.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brunn G. J., Hudson C. C., Sekulić A., Williams J. M., Hosoi H., Houghton P. J., Lawrence J. C., Jr, Abraham R. T. Phosphorylation of the translational repressor PHAS-I by the mammalian target of rapamycin. Science. 1997 Jul 4;277(5322):99–101. doi: 10.1126/science.277.5322.99. [DOI] [PubMed] [Google Scholar]
  6. Burnett P. E., Barrow R. K., Cohen N. A., Snyder S. H., Sabatini D. M. RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1432–1437. doi: 10.1073/pnas.95.4.1432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cafferkey R., McLaughlin M. M., Young P. R., Johnson R. K., Livi G. P. Yeast TOR (DRR) proteins: amino-acid sequence alignment and identification of structural motifs. Gene. 1994 Apr 8;141(1):133–136. doi: 10.1016/0378-1119(94)90141-4. [DOI] [PubMed] [Google Scholar]
  8. Chen J., Peterson R. T., Schreiber S. L. Alpha 4 associates with protein phosphatases 2A, 4, and 6. Biochem Biophys Res Commun. 1998 Jun 29;247(3):827–832. doi: 10.1006/bbrc.1998.8792. [DOI] [PubMed] [Google Scholar]
  9. Di Como C. J., Arndt K. T. Nutrients, via the Tor proteins, stimulate the association of Tap42 with type 2A phosphatases. Genes Dev. 1996 Aug 1;10(15):1904–1916. doi: 10.1101/gad.10.15.1904. [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. 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. Hara K., Yonezawa K., Weng Q. P., Kozlowski M. T., Belham C., Avruch J. Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism. J Biol Chem. 1998 Jun 5;273(23):14484–14494. doi: 10.1074/jbc.273.23.14484. [DOI] [PubMed] [Google Scholar]
  14. Hein C., Springael J. Y., Volland C., Haguenauer-Tsapis R., André B. NPl1, an essential yeast gene involved in induced degradation of Gap1 and Fur4 permeases, encodes the Rsp5 ubiquitin-protein ligase. Mol Microbiol. 1995 Oct;18(1):77–87. doi: 10.1111/j.1365-2958.1995.mmi_18010077.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Heitman J., Movva N. R., Hall M. N. Targets for cell cycle arrest by the immunosuppressant rapamycin in yeast. Science. 1991 Aug 23;253(5022):905–909. doi: 10.1126/science.1715094. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Helliwell S. B., Howald I., Barbet N., Hall M. N. TOR2 is part of two related signaling pathways coordinating cell growth in Saccharomyces cerevisiae. Genetics. 1998 Jan;148(1):99–112. doi: 10.1093/genetics/148.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Helliwell S. B., Wagner P., Kunz J., Deuter-Reinhard M., Henriquez R., Hall M. N. TOR1 and TOR2 are structurally and functionally similar but not identical phosphatidylinositol kinase homologues in yeast. Mol Biol Cell. 1994 Jan;5(1):105–118. doi: 10.1091/mbc.5.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hicke L., Riezman H. Ubiquitination of a yeast plasma membrane receptor signals its ligand-stimulated endocytosis. Cell. 1996 Jan 26;84(2):277–287. doi: 10.1016/s0092-8674(00)80982-4. [DOI] [PubMed] [Google Scholar]
  21. Hicke L. Ubiquitin-dependent internalization and down-regulation of plasma membrane proteins. FASEB J. 1997 Dec;11(14):1215–1226. doi: 10.1096/fasebj.11.14.9409540. [DOI] [PubMed] [Google Scholar]
  22. Hoffman C. S., Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. doi: 10.1016/0378-1119(87)90131-4. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Keith C. T., Schreiber S. L. PIK-related kinases: DNA repair, recombination, and cell cycle checkpoints. Science. 1995 Oct 6;270(5233):50–51. doi: 10.1126/science.270.5233.50. [DOI] [PubMed] [Google Scholar]
  25. Koller A., Heitman J., Hall M. N. Regional bivalent-univalent pairing versus trivalent pairing of a trisomic chromosome in Saccharomyces cerevisiae. Genetics. 1996 Nov;144(3):957–966. doi: 10.1093/genetics/144.3.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kunz J., Henriquez R., Schneider U., Deuter-Reinhard M., Movva N. R., Hall M. N. Target of rapamycin in yeast, TOR2, is an essential phosphatidylinositol kinase homolog required for G1 progression. Cell. 1993 May 7;73(3):585–596. doi: 10.1016/0092-8674(93)90144-f. [DOI] [PubMed] [Google Scholar]
  27. Lane D. Awakening angels. Nature. 1998 Aug 13;394(6694):616–617. doi: 10.1038/29166. [DOI] [PubMed] [Google Scholar]
  28. Marchal C., Haguenauer-Tsapis R., Urban-Grimal D. A PEST-like sequence mediates phosphorylation and efficient ubiquitination of yeast uracil permease. Mol Cell Biol. 1998 Jan;18(1):314–321. doi: 10.1128/mcb.18.1.314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Murata K., Wu J., Brautigan D. L. B cell receptor-associated protein alpha4 displays rapamycin-sensitive binding directly to the catalytic subunit of protein phosphatase 2A. Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10624–10629. doi: 10.1073/pnas.94.20.10624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Noda T., Ohsumi Y. Tor, a phosphatidylinositol kinase homologue, controls autophagy in yeast. J Biol Chem. 1998 Feb 13;273(7):3963–3966. doi: 10.1074/jbc.273.7.3963. [DOI] [PubMed] [Google Scholar]
  31. Pullen N., Thomas G. The modular phosphorylation and activation of p70s6k. FEBS Lett. 1997 Jun 23;410(1):78–82. doi: 10.1016/s0014-5793(97)00323-2. [DOI] [PubMed] [Google Scholar]
  32. Schmidt A., Bickle M., Beck T., Hall M. N. The yeast phosphatidylinositol kinase homolog TOR2 activates RHO1 and RHO2 via the exchange factor ROM2. Cell. 1997 Feb 21;88(4):531–542. doi: 10.1016/s0092-8674(00)81893-0. [DOI] [PubMed] [Google Scholar]
  33. Schmidt A., Hall M. N., Koller A. Two FK506 resistance-conferring genes in Saccharomyces cerevisiae, TAT1 and TAT2, encode amino acid permeases mediating tyrosine and tryptophan uptake. Mol Cell Biol. 1994 Oct;14(10):6597–6606. doi: 10.1128/mcb.14.10.6597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schmidt A., Kunz J., Hall M. N. TOR2 is required for organization of the actin cytoskeleton in yeast. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13780–13785. doi: 10.1073/pnas.93.24.13780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sophianopoulou V., Diallinas G. Amino acid transporters of lower eukaryotes: regulation, structure and topogenesis. FEMS Microbiol Rev. 1995 Jan;16(1):53–75. doi: 10.1111/j.1574-6976.1995.tb00155.x. [DOI] [PubMed] [Google Scholar]
  36. Springael J. Y., André B. Nitrogen-regulated ubiquitination of the Gap1 permease of Saccharomyces cerevisiae. Mol Biol Cell. 1998 Jun;9(6):1253–1263. doi: 10.1091/mbc.9.6.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stan R., McLaughlin M. M., Cafferkey R., Johnson R. K., Rosenberg M., Livi G. P. Interaction between FKBP12-rapamycin and TOR involves a conserved serine residue. J Biol Chem. 1994 Dec 23;269(51):32027–32030. [PubMed] [Google Scholar]
  38. Stanbrough M., Magasanik B. Transcriptional and posttranslational regulation of the general amino acid permease of Saccharomyces cerevisiae. J Bacteriol. 1995 Jan;177(1):94–102. doi: 10.1128/jb.177.1.94-102.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Thomas G., Hall M. N. TOR signalling and control of cell growth. Curr Opin Cell Biol. 1997 Dec;9(6):782–787. doi: 10.1016/s0955-0674(97)80078-6. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. Zheng X. F., Florentino D., Chen J., Crabtree G. R., Schreiber S. L. TOR kinase domains are required for two distinct functions, only one of which is inhibited by rapamycin. Cell. 1995 Jul 14;82(1):121–130. doi: 10.1016/0092-8674(95)90058-6. [DOI] [PubMed] [Google Scholar]
  43. von Manteuffel S. R., Gingras A. C., Ming X. F., Sonenberg N., Thomas G. 4E-BP1 phosphorylation is mediated by the FRAP-p70s6k pathway and is independent of mitogen-activated protein kinase. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4076–4080. doi: 10.1073/pnas.93.9.4076. [DOI] [PMC free article] [PubMed] [Google Scholar]

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