Two novel targets of the MAP kinase Kss1 are negative regulators of invasive growth in the yeast Saccharomyces cerevisiae.
10.1101/gad.10.22.2831
Crossref journal-article
Cold Spring Harbor Laboratory
Genes & Development (246)
Abstract

Haploid cells of budding yeast Saccharomyces cerevisiae respond to mating pheromones by inducing genes required for conjugation, arresting cell cycle progression, and undergoing morphological changes. The same cells respond to nutrient deprivation by altering budding pattern and inducing genes required for invasive growth. Both developmental alternatives to vegetative proliferation require the MAP kinase Kss1 and the transcriptional transactivator Ste12. Using a two-hybrid screen for gene products that interact with Kss1, two homologous and previously uncharacterized loci (DIG1 and DIG2, for down-regulator of invasive growth) were identified. DIG2 is pheromone-inducible, whereas DIG1 is constitutively expressed. Dig1 colocalizes with Kssl in the nucleus, coimmunoprecipitates with Kss1 from cell extracts in a pheromone-independent manner, and is phosphorylated by Kss1 in immune complexes in a pheromone-stimulated manner. Kss1 binds specifically to a GST-Dig1 fusion in the absence of any other yeast protein. Using the two-hybrid method, both Dig1 and Dig2 also interact with the other MAP kinase of the pheromone response pathway, Fus3. However, neither dig1 or dig2 single mutants, nor a dig1 dig2 double mutant, have a discernible effect on mating. In contrast, dig1 dig2 cells constitutively invade agar medium, whereas a dig1 dig2 ste12 triple mutant does not, indicating that Dig1 and Dig2 share a role in negatively regulating the invasive growth pathway. High-level expression of Dig1 suppresses invasive growth and also causes cells to appear more resistant to pheromone-imposed cell cycle arrest. Ste12 also binds specifically to GST-Dig1 in the absence of any other yeast protein. Collectively, these findings indicate that Dig1, and most likely Dig2, are physiological substrates of Kssl and suggest that they regulate Ste12 function by direct protein-protein interaction.

Bibliography

Cook, J. G., Bardwell, L., Kron, S. J., & Thorner, J. (1996). Two novel targets of the MAP kinase Kss1 are negative regulators of invasive growth in the yeast Saccharomyces cerevisiae. Genes & Development, 10(22), 2831–2848.

Authors 4
  1. J G Cook (first)
  2. L Bardwell (additional)
  3. S J Kron (additional)
  4. J Thorner (additional)
References 98 Referenced 189
  1. 10.1006/jmbi.1990.9999
  2. 10.1016/S0021-9258(18)98613-8 / J. Biol. Chem. / Pro-Leu-Ser/ Thr-Pro is a consensus primary sequence for substrate protein phosphorylation. Characterization of the phosphorylation of c-myc and c-jun proteins by an epidermal growth factor receptor threonine 669 protein kinase. (1991)
  3. 10.1073/pnas.91.9.3926
  4. 10.1006/dbio.1994.1323
  5. 10.1128/MCB.16.7.3637 / Mol. Cell. Biol. / Signaling in the yeast pheromone response pathway: Specific and high-affinity interaction of the mitogenactivated protein (MAP) kinases Kss1 and Fus3 with the upstream MAP kinase kinase Ste7. (1996)
  6. 10.1016/0076-6879(95)54018-0
  7. {'key': '2021111418410728000_10.22.2831.7', 'first-page': '353', 'article-title': 'Signal-mediated import of bacteriophage T7 RNA polymerase into the Saccharomyces cerevisiae nucleus and specific transcription of target genes.', 'volume': '10', 'year': '1990', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Signal-mediated import of bacteriophage T7 RNA polymerase into the Saccharomyces cerevisiae nucleus and specific transcription of target genes. (1990)
  8. 10.1083/jcb.127.3.623
  9. {'key': '2021111418410728000_10.22.2831.9', 'first-page': '5567', 'article-title': 'Regulation of dimorphism in Saccharomyces cerevisiae: Involvement of the novel protein kinase homolog Elmlp and protein phosphatase 2A.', 'volume': '13', 'year': '1993', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Regulation of dimorphism in Saccharomyces cerevisiae: Involvement of the novel protein kinase homolog Elmlp and protein phosphatase 2A. (1993)
  10. 10.1093/genetics/140.4.1259 / Genetics / Mutational analysis of morphologic differentiation in Saccharomyces cerevisiae. (1995)
  11. 10.1007/BF00330984
  12. 10.1016/0003-2697(76)90527-3
  13. 10.1101/SQB.1985.050.01.078
  14. 10.1101/gad.6.7.1305
  15. 10.1128/jb.155.2.903-906.1983 / J. Bacteriol. / Extracellular suppression allows mating by pheromone-deficient sterile mutants of Saccharomyces cerevisiae. (1983)
  16. 10.1016/0092-8674(90)90503-7
  17. {'key': '2021111418410728000_10.22.2831.17', 'first-page': '445', 'article-title': 'Phosphorylation of FAR1 in response to alpha-factor: A possible requirement for cell-cycle arrest.', 'volume': '4', 'year': '1992', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Phosphorylation of FAR1 in response to alpha-factor: A possible requirement for cell-cycle arrest. (1992)
  18. 10.1016/S0092-8674(00)80972-1
  19. 10.1016/0092-8674(94)90427-8
  20. 10.1016/0092-8674(79)90117-X
  21. 10.1016/S0021-9258(18)98601-1 / J. Biol. Chem. / Definition of a consensus sequence for peptide substrate recognition by p44mpk, the meiosis-activated myelin basic protein kinase. (1991)
  22. 10.1016/0092-8674(89)90509-6
  23. 10.1016/S0021-9258(18)82362-6 / J. Biol. Chem. / The mitogen-activated protein kinase signal transduction pathway. (1993)
  24. 10.1016/0092-8674(86)90599-4
  25. 10.1210/edrv-16-3-259
  26. {'key': '2021111418410728000_10.22.2831.26', 'first-page': '4169', 'article-title': 'Pheromonal regulation and sequence of the Saccharomyes cerevisiae SST2 gene: A model for desensitization to pheromone.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Pheromonal regulation and sequence of the Saccharomyes cerevisiae SST2 gene: A model for desensitization to pheromone. (1987)
  27. 10.1128/MCB.16.9.5194 / Mol. Cell. Biol. / Sst, a negative regulator of pheromone signaling in the yeast Saccharomyces cerevisiae: Expression, localization, genetic interaction and physical association with Gpal (G protein a subunit). (1996)
  28. 10.1073/pnas.86.15.5703
  29. 10.1101/gad.7.4.555
  30. 10.1016/0092-8674(90)90668-5
  31. 10.1073/pnas.88.21.9392
  32. 10.1091/mbc.4.5.495
  33. 10.1098/rstb.1996.0010 / Philos. Trans. R. Soc. Lond. B. Biol. Sci. / Feedback regulation of MAP kinase signal pathways. (1996)
  34. 10.1038/362261a0
  35. {'key': '2021111418410728000_10.22.2831.35', 'first-page': '3610', 'article-title': 'Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.', 'volume': '5', 'year': '1985', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. (1985)
  36. 10.1016/0003-2697(84)90381-6
  37. 10.1038/340245a0
  38. 10.1101/gad.6.7.1280
  39. 10.1111/j.1365-2958.1996.tb02470.x
  40. 10.1016/0378-1119(88)90185-0
  41. 10.1016/0092-8674(92)90079-R
  42. 10.1111/j.1432-1033.1990.tb19384.x
  43. 10.1016/S0022-2836(83)80284-8
  44. Harlow, E. and D. Lane. 1988. Antibodies: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
  45. {'key': '2021111418410728000_10.22.2831.45', 'first-page': '5767', 'article-title': 'CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: Identification, characterization, and homology to the B sub-unit of mammalian type 2A protein phosphatase.', 'volume': '11', 'year': '1991', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: Identification, characterization, and homology to the B sub-unit of mammalian type 2A protein phosphatase. (1991)
  46. 10.1016/0076-6879(91)94011-Z
  47. Herskowitz, I., J. Rine, and J. Strthern. 1992. Mating-type determination and mating-type interconversion in Saccharomyces cerevisiae. In The molecular and cellular biology of the yeast Saccharomyces: Gene expression (ed. E.W. Jones, J.R. Pringle, and J.R. Broach), pp. 583–656. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  48. 10.1002/yea.320020304
  49. 10.1016/S0021-9258(19)70246-4 / J. Biol. Chem. / Isolation and characterization of the yeast 3-phosphoglycerokinase gene (PGK1) by an immunological screening technique. (1980)
  50. James, P., Halladay, J. and Craig. E.A. 1996. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics (in press). (10.1093/genetics/144.4.1425)
  51. 10.1016/0076-6879(91)94034-A / Methods Enymol. / Tackling the protease problem in yeast. (1991)
  52. 10.1002/yea.320060502
  53. 10.1016/0092-8674(83)90070-3
  54. 10.1016/0003-9861(65)90096-2
  55. 10.1016/0076-6879(91)94030-G
  56. 10.1101/gad.8.3.313
  57. 10.1016/0955-0674(95)80069-7
  58. {'key': '2021111418410728000_10.22.2831.58', 'first-page': '003', 'article-title': 'Symmetric cell division in pseudohyphae of the yeast Saccharomyces cerevisiae.', 'volume': '51', 'year': '1994', 'journal-title': 'Mol. Biol. Cell'} / Mol. Biol. Cell / Symmetric cell division in pseudohyphae of the yeast Saccharomyces cerevisiae. (1994)
  59. 10.1146/annurev.ge.27.120193.001051
  60. 10.1093/nar/22.6.999
  61. 10.1002/j.1460-2075.1992.tb05587.x / EMBO J. / The protein kinase homolog Ste20p is required to link the yeast pheromone response G-protein βγ subunits to downstream signalling components. (1992)
  62. 10.1126/science.270.5239.1210
  63. 10.1126/science.2983426
  64. 10.1093/genetics/132.3.665 / Genetics / Construction of a Gal1-regulated yeast cDNA expression library and its application to the identification of genes whose overexpression causes lethality in yeast. (1992)
  65. 10.1126/science.8259520
  66. 10.1016/0378-1119(95)00144-U
  67. 10.1091/mbc.6.7.889
  68. 10.1038/367040a0
  69. 10.1073/pnas.91.16.7762
  70. 10.1002/j.1460-2075.1995.tb07189.x / EMBO J. / A novel serine kinase activated by racl/CDC42Hs-dependent autophosphorylation is related to PAK65 and STE20. (1995)
  71. Miller, J.H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
  72. 10.1073/pnas.93.11.5352
  73. 10.1073/pnas.91.8.3398
  74. 10.1002/yea.320030303
  75. 10.1146/annurev.bi.62.070193.001251
  76. 10.1016/0092-8674(93)90254-N
  77. 10.1093/genetics/138.3.609 / Genetics / Protein-protein interactions in the yeast pheromone response pathway: Ste5P interacts with all members of the MAP kinase cascade. (1994)
  78. 10.1016/0092-8674(95)90006-3
  79. 10.1016/0092-8674(88)90045-1
  80. 10.1101/gad.8.24.2974
  81. Sambrook, J., E. Fritsch, and T. Maniatis. 1989. Molecular cloning: A laboratory manual, 2nd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  82. 10.1073/pnas.76.10.4951
  83. Sherman, F., G.R. Fink, and J.B. Hicks. 1986. Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
  84. 10.1093/genetics/122.1.19 / Genetics / A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. (1989)
  85. 10.1038/376702a0
  86. 10.1016/0022-2836(75)90217-X
  87. 10.1016/0076-6879(91)94008-Z
  88. Sprague, G.F., Jr. and J. Thorner. 1992. Pheromone response and signal transduction during the mating process of Saccharomyces cerevisiae. In The molecular and cellular biology of the yeast Saccharomyces: Gene expression (ed. E.W. Jones, J.R. Pringle, and J.R. Broach), pp. 657–744. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  89. 10.1016/0022-2836(91)90855-Z
  90. 10.1042/bj2880351
  91. 10.1016/S0960-9822(00)00154-8
  92. 10.1128/MCB.13.9.5659
  93. 10.1073/pnas.90.13.6213
  94. Van Arsdell, S.W. and J. Thorner. 1987. Hormonal regulation of gene expression in yeast. In Transcriptional control mechanisms (ed. D.K. Granner, M.G. Rosenfeld, and S. Chang), pp. 325–332. Alan R. Liss, New York, NY.
  95. 10.1016/0092-8674(89)90249-3
  96. 10.1016/S0065-2911(08)60394-X
  97. 10.1074/jbc.270.27.15984
  98. 10.1128/MCB.15.10.5246 / Mol. Cell. Biol. / Pheromone signalling in Saccharomyces cerevisiae requires the small GTP-binding protein Cdc42p and its activator Cdc24. (1995)
Dates
Type When
Created 18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited 3 years, 9 months ago (Nov. 14, 2021, 9:55 p.m.)
Indexed 11 months, 2 weeks ago (Sept. 19, 2024, 11:09 a.m.)
Issued 28 years, 9 months ago (Nov. 15, 1996)
Published 28 years, 9 months ago (Nov. 15, 1996)
Published Online 28 years, 9 months ago (Nov. 15, 1996)
Published Print 28 years, 9 months ago (Nov. 15, 1996)
Funders 0

None

@article{Cook_1996, title={Two novel targets of the MAP kinase Kss1 are negative regulators of invasive growth in the yeast Saccharomyces cerevisiae.}, volume={10}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.10.22.2831}, DOI={10.1101/gad.10.22.2831}, number={22}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Cook, J G and Bardwell, L and Kron, S J and Thorner, J}, year={1996}, month=nov, pages={2831–2848} }