Protein-Protein Interactions in the Yeast Pheromone Response Pathway: Ste5p Interacts With All Members of the MAP Kinase Cascade

INVESTIGATIONS

Protein-Protein Interactions in the Yeast Pheromone Response Pathway: Ste5p Interacts With All Members of the MAP Kinase Cascade

J. A. Printen and G. F. Sprague-Jr.
Institute of Molecular Biology and Departments of Chemistry, University of Oregon, Eugene, Oregon 97403-1229

We have used the two-hybrid system of Fields and Song to identify protein-protein interactions that occur in the pheromone response pathway of the yeast Saccharomyces cerevisiae. Pathway components Ste4p, Ste5p, Ste7p, Ste11p, Ste12p, Ste20p, Fus3p and Kss1p were tested in all pairwise combinations. All of the interactions we detected involved at least one member of the MAP kinase cascade that is a central element of the response pathway. Ste5p, a protein of unknown biochemical function, interacted with protein kinases that operate at each step of the MAP kinase cascade, specifically with Ste11p (an MEKK), Ste7p (an MEK), and Fus3p (a MAP kinase). This finding suggests that one role of Ste5p is to serve as a scaffold to facilitate interactions among members of the kinase cascade. In this role as facilitator, Ste5p may make both signal propagation and signal attenuation more efficient. Ste5p may also help minimize cross-talk with other MAP kinase cascades and thus ensure the integrity of the pheromone response pathway. We also found that both Ste11p and Ste7p interact with Fus3p and Kss1p. Finally, we detected an interaction between one of the MAP kinases, Kss1p, and a presumptive target, the transcription factor Ste12p. We failed to detect interactions of Ste4p or Ste20p with any other component of the response pathway.

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				J. M.-Y. Lu, R. J. Deschenes, and J. S. Fassler Role for the Ran Binding Protein, Mog1p, in Saccharomyces cerevisiae SLN1-SKN7 Signal Transduction Eukaryot. Cell, December 1, 2004; 3(6): 1544 - 1556. [Abstract] [Full Text] [PDF]

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				S.-H. Park, A. Zarrinpar, and W. A. Lim Rewiring MAP Kinase Pathways Using Alternative Scaffold Assembly Mechanisms Science, February 14, 2003; 299(5609): 1061 - 1064. [Abstract] [Full Text] [PDF]

				C. Ruiz, V. Escribano, E. Morgado, M. Molina, and M. J. Mazon Cell-type-dependent repression of yeast a-specific genes requires Itc1p, a subunit of the Isw2p-Itc1p chromatin remodelling complex Microbiology, February 1, 2003; 149(2): 341 - 351. [Abstract] [Full Text] [PDF]

				R. K. Esch and B. Errede Pheromone induction promotes Ste11 degradation through a MAPK feedback and ubiquitin-dependent mechanism PNAS, July 9, 2002; 99(14): 9160 - 9165. [Abstract] [Full Text] [PDF]

				G. R. Smith, S. A. Givan, P. Cullen, and G. F. Sprague Jr. GTPase-Activating Proteins for Cdc42 Eukaryot. Cell, June 1, 2002; 1(3): 469 - 480. [Abstract] [Full Text] [PDF]

				E. A. Elion The Ste5p scaffold J. Cell Sci., March 13, 2002; 114(22): 3967 - 3978. [Abstract] [Full Text] [PDF]

				J. R. Morris, N. H. Keep, and E. Solomon Identification of Residues Required for the Interaction of BARD1 with BRCA1 J. Biol. Chem., March 8, 2002; 277(11): 9382 - 9386. [Abstract] [Full Text] [PDF]

				Z.-x. Zhou, B. He, S. H. Hall, E. M. Wilson, and F. S. French Domain Interactions between Coregulator ARA70 and the Androgen Receptor (AR) Mol. Endocrinol., February 1, 2002; 16(2): 287 - 300. [Abstract] [Full Text] [PDF]

				S. Erdman and M. Snyder A Filamentous Growth Response Mediated by the Yeast Mating Pathway Genetics, November 1, 2001; 159(3): 919 - 928. [Abstract] [Full Text] [PDF]