14-3-3 proteins interact with specific MEK kinases

Gary R. Fanger, Christian Widmann, Amy C. Porter, Sue Sather, Gary L. Johnson, Richard Vaillancourt

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

MEK (mitogen-activated protein kinase/extracellular signal-regulated kinase kinase) kinases (MEKKs) regulate c-Jun N-terminal kinase and extracellular response kinase pathways. The 14-3-3ζ and 14-3-3ε isoforms were isolated in a two-hybrid screen for proteins interacting with the N- terminal regulatory domain of MEKK3. 14-3-3 proteins bound both the N- terminal regulatory and C-terminal kinase domains of MEKK3. The binding affinity of 14-3-3 for the MEKK3 N terminus was 90 nM, demonstrating a high affinity interaction. 14-3-3 proteins also interacted with MEKK1 and MEKK2, but not MEKK4. Endogenous 14-3-3 protein and MEKK1 and MEKK2 were similarly distributed in the cell, consistent with their in vitro interactions. MEKK1 and 14-3-3 proteins colocalized using two-color digital confocal immunofluorescence. Binding of 14-3-3 proteins mapped to the N-terminal 393 residues of 196-kDa MEKK1. Unlike MEKK2 and MEKK3, the C-terminal kinase domain of MEKK1 demonstrated little or no ability to interact with 14-3-3 proteins. MEKK1, but not MEKK2, -3 or -4, is a caspase-3 substrate that when cleaved releases the kinase domain from the N-terminal regulatory domain. Functionally, caspase-3 cleavage of MEKK1 releases the kinase domain from the N-terminal 14-3-3-binding region, demonstrating that caspases can selectively alter protein kinase interactions with regulatory proteins. With regard to MEKK1, -2 and -3, 14-3-3 proteins do not appear to directly influence activity, but rather function as 'scaffolds' for protein-protein interactions.

Original languageEnglish (US)
Pages (from-to)3476-3483
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number6
DOIs
StatePublished - Feb 6 1998

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14-3-3 Proteins
MAP Kinase Kinase Kinases
Phosphotransferases
Caspase 3
Proteins
JNK Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinase Kinases
Extracellular Signal-Regulated MAP Kinases
Caspases
Mitogen-Activated Protein Kinases
Scaffolds
Protein Kinases
Fluorescent Antibody Technique
Protein Isoforms
Color
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fanger, G. R., Widmann, C., Porter, A. C., Sather, S., Johnson, G. L., & Vaillancourt, R. (1998). 14-3-3 proteins interact with specific MEK kinases. Journal of Biological Chemistry, 273(6), 3476-3483. https://doi.org/10.1074/jbc.273.6.3476

14-3-3 proteins interact with specific MEK kinases. / Fanger, Gary R.; Widmann, Christian; Porter, Amy C.; Sather, Sue; Johnson, Gary L.; Vaillancourt, Richard.

In: Journal of Biological Chemistry, Vol. 273, No. 6, 06.02.1998, p. 3476-3483.

Research output: Contribution to journalArticle

Fanger, GR, Widmann, C, Porter, AC, Sather, S, Johnson, GL & Vaillancourt, R 1998, '14-3-3 proteins interact with specific MEK kinases', Journal of Biological Chemistry, vol. 273, no. 6, pp. 3476-3483. https://doi.org/10.1074/jbc.273.6.3476
Fanger, Gary R. ; Widmann, Christian ; Porter, Amy C. ; Sather, Sue ; Johnson, Gary L. ; Vaillancourt, Richard. / 14-3-3 proteins interact with specific MEK kinases. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 6. pp. 3476-3483.
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