A Conserved Docking Site in MEKs Mediates High-affinity Binding to MAP Kinases and Cooperates with a Scaffold Protein to Enhance Signal Transmission

A. Jane Bardwell, Laura J. Flatauer, Karen Matsukuma, Jeremy Thorner, Lee Bardwell

Research output: Contribution to journalArticle

140 Citations (Scopus)

Abstract

The recognition of mitogen-activated protein kinases (MAPKs) by their upstream activators, MAPK/ERK kinases (MEKs), is crucial for the effective and accurate transmission of many signals. We demonstrated previously that the yeast MAPKs Kss1 and Fus3 bind with high affinity to the N terminus of the MEK Ste7, and proposed that a conserved motif in Ste7, the MAPK-docking site, mediates this interaction. Here we show that the corresponding sequences in human MEK1 and MEK2 are necessary and sufficient for the direct binding of the MAPKs ERK1 and ERK2. Mutations in MEK1, MEK2, or Ste7 that altered conserved residues in the docking site diminished binding of the cognate MAPKs. Furthermore, short peptides corresponding to the docking sites in these MEKs inhibited MEK1-mediated phosphorylation of ERK2 in vitro. In yeast cells, docking-defective alleles of Ste7 were modestly compromised in their ability to transmit the mating pheromone signal. This deficiency was dramatically enhanced when the ability of the Ste5 scaffold protein to associate with components of the MAPK cascade was also compromised. Thus, both the MEK-MAPK docking interaction and binding to the Ste5 scaffold make mutually reinforcing contributions to the efficiency of signaling by this MAPK cascade in vivo.

Original languageEnglish (US)
Pages (from-to)10374-10386
Number of pages13
JournalJournal of Biological Chemistry
Volume276
Issue number13
DOIs
StatePublished - Mar 30 2001
Externally publishedYes

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Mitogen-Activated Protein Kinase Kinases
Mitogen-Activated Protein Kinases
Scaffolds
Phosphotransferases
Proteins
Yeast
Yeasts
Pheromones
Phosphorylation
Alleles
Binding Sites
Cells
Peptides
Mutation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A Conserved Docking Site in MEKs Mediates High-affinity Binding to MAP Kinases and Cooperates with a Scaffold Protein to Enhance Signal Transmission. / Jane Bardwell, A.; Flatauer, Laura J.; Matsukuma, Karen; Thorner, Jeremy; Bardwell, Lee.

In: Journal of Biological Chemistry, Vol. 276, No. 13, 30.03.2001, p. 10374-10386.

Research output: Contribution to journalArticle

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