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Updated: Jan 6, 2026

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells
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Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells

Published on: March 1, 2024

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Architecture of autoinhibited and active BRAF-MEK1-14-3-3 complexes

Eunyoung Park1,2, Shaun Rawson2, Kunhua Li1,2

  • 1Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA.

Nature
|October 4, 2019
PubMed

Insights

Structural insights reveal how 14-3-3 proteins regulate BRAF kinase activity. These findings explain BRAF

Area of Science:

  • Molecular Biology
  • Structural Biology
  • Biochemistry

Background:

  • RAF kinases, including BRAF, are key regulators of the MAP kinase cascade controlling cell growth.
  • Dysregulated RAF activity is implicated in various cancers, yet its structural regulation remains unclear.
  • Understanding BRAF regulation is crucial for developing targeted cancer therapies.

Purpose of the Study:

  • To elucidate the structural mechanisms underlying BRAF kinase autoinhibition and activation.
  • To investigate the role of 14-3-3 proteins in BRAF regulation.
  • To provide a structural basis for understanding BRAF mutations in cancer.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) was employed to determine high-resolution structures.
  • Structures of full-length BRAF were solved in complex with MEK1 and 14-3-3 dimers.
  • Analysis focused on autoinhibited and active states of BRAF.

Main Results:

  • An autoinhibited BRAF-MEK1 complex is stabilized by a 14-3-3 dimer binding to phosphorylated BRAF sites.
  • 14-3-3 binding sequesters key BRAF domains, preventing kinase dimerization and activity.
  • Activation involves 14-3-3 mediating the formation of an active, dimeric BRAF kinase.

Conclusions:

  • The 14-3-3 dimer acts as a crucial scaffold, controlling BRAF kinase autoinhibition and activation.
  • These structural findings offer insights into BRAF regulation and its aberrant function in disease.
  • The study provides a framework for understanding BRAF-related cancers and developmental disorders.

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