Hydrophobic as Well as Charged Residues in Both MEK1 and ERK2 Are Important for Their Proper Docking
Open accessAbstract
Docking between MEK1 and ERK2 is required for their stable interaction and efficient signal transmission. The MEK1 N terminus contains the ERK docking or D domain that consists of conserved hydrophobic and basic residues. We mutated the hydrophobic and basic residues individually and found that loss of either type reduced MEK1 phosphorylation of ERK2 in vitro and its ability to bind to ERK2 in vivo. Moreover, ERK2 was localized in both the cytoplasm and the nucleus when co-expressed with MEK1 that had mutations in either the hydrophobic or the basic residues. We then identified two conserved hydrophobic residues on ERK2 that play roles in docking with MEK1. Mutating these residues to alanine reduced the interaction of ERK2 with MEK1 in cells. These mutations also reduced the phosphorylation of MEK1 by ERK2 but had little effect on phosphorylation of MBP by ERK2. Finally, we generated docking site mutants in ERK2-MEK1 fusion proteins. Although the mutation of the MEK1 D domain significantly reduced ERK2-MEK1 activity, mutations of the putatively complementary acidic residues and hydrophobic residues on ERK2 did not change its activity. However, both types of mutations decreased the phosphorylation of Elk-1 caused by ERK2-MEK1 fusion proteins. These findings suggest complex interactions of MEK1 D domains with ERK2 that influence its activation and its effects on substrates.
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MKKs MEK1/2, MKK3/6, MKK4/7, and STE7 possess a flexible N-terminal extension that includes the 15 amino acid KIM, followed by their dual-specificity kinase domain.
“For example, the MKKs MEK1/2, MKK3/6, MKK4/7, and STE7, representing the MAP2Ks of ERK, p38, JNK, and FUS3, respectively, possess a flexible Nterminal extension, which includes the 15 amino acid KIM, followed by their dual-specificity kinase domain [Fig.1(B), left].”
From Molecular basis of MAP kinase regulation (2013) · cites this paper as [23]
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- Authors
- Xu, B.-E., Stippec, S., Robinson, F. L., & Cobb, M. H.
- Affiliations
- The University of Texas Southwestern Medical Center (US)
- Corresponding author
- Cobb, Melanie H.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2001-07
- Volume
- 276
- Issue
- 28
- Pages
- 26509-26515
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1973473002
- Cited by
- 91
- References
- 27
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 2 · 2023: 1 · 2022: 3 · 2019: 2 · 2017: 2 · 2016: 1 · 2015: 1 · 2014: 2 · 2013: 5 · 2012: 1
- FWCI
- 3.85
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-34128)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Keywords
- MEK1, ERK2, D domain, hydrophobic residues, basic residues, protein-protein interaction, MEK1 phosphorylation, ERK2 phosphorylation, Elk-1 phosphorylation, MAPK signaling
- MeSH terms
- Amino Acid Sequence, Cell Line, Humans, Molecular Sequence Data, Mutation, Structure-Activity Relationship, Signal Transduction, Protein Serine-Threonine Kinases, Mitogen-Activated Protein Kinase 1, Mitogen-Activated Protein Kinase Kinases, MAP Kinase Kinase 1
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