Activation of MEK Family Kinases Requires Phosphorylation of Two Conserved Ser/Thr Residues

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1994 Mar 1

PMID 8131746

Citations 107

Authors

C F Zheng

K L Guan

Affiliations

Soon will be listed here.

Abstract

MEK is a family of dual specific protein kinases which activate the extracellular signal-regulated kinases by phosphorylation of threonine and tyrosine residues. MEK itself is activated via serine phosphorylation by upstream activator kinases, including c-raf, mos and MEK kinase. Here, we report the activation phosphorylation sites of human MEK1 and yeast STE7 kinase as determined by a combination of biochemical and genetic approaches. In human MEK1, substitution of either serine residue 218 or 222 with alanine completely abolished its activation by epidermal growth factor-stimulated Swiss 3T3 cell lysates or immunoprecipitated c-raf, suggesting that both serine residues are required for MEK1 activation. Phosphopeptide analysis demonstrated that serine residues 218 and 222 of human MEK1 are the primary sites for phosphorylation by c-raf. These two serine residues are highly conserved in all members of the MEK family, including the yeast STE7 gene product, a MEK homolog in the yeast mating pheromone response pathway. Mutation of the corresponding residues in STE7 completely abolished the biological functions of this gene. These data demonstrate that MEK is activated by phosphorylation of two adjacent serine/threonine residues and this activation mechanism is conserved in the MEK family kinases.

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References

Tsuda L, Inoue Y, Yoo M, Mizuno M, Hata M, Lim Y . A protein kinase similar to MAP kinase activator acts downstream of the raf kinase in Drosophila. Cell. 1993; 72(3):407-14. DOI: 10.1016/0092-8674(93)90117-9. View

Zheng C, Guan K . Properties of MEKs, the kinases that phosphorylate and activate the extracellular signal-regulated kinases. J Biol Chem. 1993; 268(32):23933-9. View

Ashworth A, Nakielny S, Cohen P, Marshall C . The amino acid sequence of a mammalian MAP kinase kinase. Oncogene. 1992; 7(12):2555-6. View

Seger R, Seger D, Lozeman F, Ahn N, Graves L, Campbell J . Human T-cell mitogen-activated protein kinase kinases are related to yeast signal transduction kinases. J Biol Chem. 1992; 267(36):25628-31. View

Gomez N, Traverse S, Cohen P . Identification of a MAP kinase kinase kinase in phaeochromocytoma (PC12) cells. FEBS Lett. 1992; 314(3):461-5. DOI: 10.1016/0014-5793(92)81527-s. View

Wu J, Harrison J, Vincent L, Haystead C, Haystead T, Michel H . Molecular structure of a protein-tyrosine/threonine kinase activating p42 mitogen-activated protein (MAP) kinase: MAP kinase kinase. Proc Natl Acad Sci U S A. 1993; 90(1):173-7. PMC: 45622. DOI: 10.1073/pnas.90.1.173. View

Ahn N, Seger R, KREBS E . The mitogen-activated protein kinase activator. Curr Opin Cell Biol. 1992; 4(6):992-9. DOI: 10.1016/0955-0674(92)90131-u. View

Matsuda S, Gotoh Y, Nishida E . Phosphorylation of Xenopus mitogen-activated protein (MAP) kinase kinase by MAP kinase kinase kinase and MAP kinase. J Biol Chem. 1993; 268(5):3277-81. View

Robbins D, Zhen E, Owaki H, Vanderbilt C, Ebert D, Geppert T . Regulation and properties of extracellular signal-regulated protein kinases 1 and 2 in vitro. J Biol Chem. 1993; 268(7):5097-106. View

10.

Posada J, Yew N, Ahn N, Woude G, Cooper J . Mos stimulates MAP kinase in Xenopus oocytes and activates a MAP kinase kinase in vitro. Mol Cell Biol. 1993; 13(4):2546-53. PMC: 359584. DOI: 10.1128/mcb.13.4.2546-2553.1993. View

11.

Brewster J, de Valoir T, Dwyer N, Winter E, Gustin M . An osmosensing signal transduction pathway in yeast. Science. 1993; 259(5102):1760-3. DOI: 10.1126/science.7681220. View

12.

Errede B, Gartner A, Zhou Z, Nasmyth K, Ammerer G . MAP kinase-related FUS3 from S. cerevisiae is activated by STE7 in vitro. Nature. 1993; 362(6417):261-4. DOI: 10.1038/362261a0. View

13.

Pleiman C, Gardner A, Blumer K, Johnson G . A divergence in the MAP kinase regulatory network defined by MEK kinase and Raf. Science. 1993; 260(5106):315-9. DOI: 10.1126/science.8385802. View

14.

Lee K, Irie K, Gotoh Y, Watanabe Y, Araki H, Nishida E . A yeast mitogen-activated protein kinase homolog (Mpk1p) mediates signalling by protein kinase C. Mol Cell Biol. 1993; 13(5):3067-75. PMC: 359699. DOI: 10.1128/mcb.13.5.3067-3075.1993. View

15.

Irie K, Takase M, Lee K, Levin D, Araki H, Matsumoto K . MKK1 and MKK2, which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol Cell Biol. 1993; 13(5):3076-83. PMC: 359700. DOI: 10.1128/mcb.13.5.3076-3083.1993. View

16.

Zheng C, Guan K . Cloning and characterization of two distinct human extracellular signal-regulated kinase activator kinases, MEK1 and MEK2. J Biol Chem. 1993; 268(15):11435-9. View

17.

Ahn N, Campbell J, Seger R, Jensen A, Graves L, KREBS E . Metabolic labeling of mitogen-activated protein kinase kinase in A431 cells demonstrates phosphorylation on serine and threonine residues. Proc Natl Acad Sci U S A. 1993; 90(11):5143-7. PMC: 46671. DOI: 10.1073/pnas.90.11.5143. View

18.

Charles C, Sun H, Lau L, Tonks N . The growth factor-inducible immediate-early gene 3CH134 encodes a protein-tyrosine-phosphatase. Proc Natl Acad Sci U S A. 1993; 90(11):5292-6. PMC: 46702. DOI: 10.1073/pnas.90.11.5292. View

19.

Errede B, Levin D . A conserved kinase cascade for MAP kinase activation in yeast. Curr Opin Cell Biol. 1993; 5(2):254-60. DOI: 10.1016/0955-0674(93)90112-4. View

20.

De Bondt H, Rosenblatt J, Jancarik J, Jones H, MORGAN D, Kim S . Crystal structure of cyclin-dependent kinase 2. Nature. 1993; 363(6430):595-602. DOI: 10.1038/363595a0. View