Functional divergence caused by mutations in an energetic hotspot in ERK2

Clinton A. Taylor, Kevin W. Cormier, Shannon E. Keenan, Svetlana Earnest, Steve Stippec, Chonlarat Wichaidit, Yu Chi Juang, Junmei Wang, Stanislav Y. Shvartsman, Elizabeth J Goldsmith, Melanie H Cobb

Research output: Contribution to journalArticle

Abstract

The most frequent extracellular signal-regulated kinase 2 (ERK2) mutation occurring in cancers is E322K (E-K). ERK2 E-K reverses a buried charge in the ERK2 common docking (CD) site, a region that binds activators, inhibitors, and substrates. Little is known about the cellular consequences associated with this mutation, other than apparent increases in tumor resistance to pathway inhibitors. ERK2 E-K, like the mutation of the preceding aspartate (ERK2 D321N [D-N]) known as the sevenmaker mutation, causes increased activity in cells and evades inactivation by dual-specificity phosphatases. As opposed to findings in cancer cells, in developmental assays in Drosophila, only ERK2 D-N displays a significant gain of function, revealing mutation-specific phenotypes. The crystal structure of ERK2 D-N is indistinguishable from that of wild-type protein, yet this mutant displays increased thermal stability. In contrast, the crystal structure of ERK2 E-K reveals profound structural changes, including disorder in the CD site and exposure of the activation loop phosphorylation sites, which likely account for the decreased thermal stability of the protein. These contiguous mutations in the CD site of ERK2 are both required for docking interactions but lead to unpredictably different functional outcomes. Our results suggest that the CD site is in an energetically strained configuration, and this helps drive conformational changes at distal sites on ERK2 during docking interactions.

Original languageEnglish (US)
Pages (from-to)15514-15523
Number of pages10
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number31
DOIs
StatePublished - Jul 30 2019

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Mitogen-Activated Protein Kinase 1
Mutation
Hot Temperature
Dual-Specificity Phosphatases
Neoplasms
Protein Stability
Mutant Proteins
Aspartic Acid
Drosophila
Phosphorylation
Phenotype

Keywords

  • ERK CD site
  • kinase
  • mutation
  • stability

ASJC Scopus subject areas

  • General

Cite this

Functional divergence caused by mutations in an energetic hotspot in ERK2. / Taylor, Clinton A.; Cormier, Kevin W.; Keenan, Shannon E.; Earnest, Svetlana; Stippec, Steve; Wichaidit, Chonlarat; Juang, Yu Chi; Wang, Junmei; Shvartsman, Stanislav Y.; Goldsmith, Elizabeth J; Cobb, Melanie H.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 31, 30.07.2019, p. 15514-15523.

Research output: Contribution to journalArticle

Taylor, CA, Cormier, KW, Keenan, SE, Earnest, S, Stippec, S, Wichaidit, C, Juang, YC, Wang, J, Shvartsman, SY, Goldsmith, EJ & Cobb, MH 2019, 'Functional divergence caused by mutations in an energetic hotspot in ERK2', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 31, pp. 15514-15523. https://doi.org/10.1073/pnas.1905015116
Taylor, Clinton A. ; Cormier, Kevin W. ; Keenan, Shannon E. ; Earnest, Svetlana ; Stippec, Steve ; Wichaidit, Chonlarat ; Juang, Yu Chi ; Wang, Junmei ; Shvartsman, Stanislav Y. ; Goldsmith, Elizabeth J ; Cobb, Melanie H. / Functional divergence caused by mutations in an energetic hotspot in ERK2. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 31. pp. 15514-15523.
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