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Cited by 10Protein Kinase Regulation and GTPase SignalingMelanoma and MAPK PathwaysProtein Tyrosine Phosphatases

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Unveiling the domain-specific and RAS isoform-specific details of BRAF kinase regulation

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Trebino, Tarah Elizabeth, Markusic, Borna, Nan, Haihan, Banerjee, Shrhea, Wang, Zhihong·2023·eLife
DOI 10.7554/elife.88836.3

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Abstract

BRAF is a key member in the MAPK signaling pathway essential for cell growth, proliferation, and differentiation. Mutant BRAF is often the underlying cause of various types of cancer and mutant RAS, the upstream regulator of BRAF, is a driver of up to one-third of all cancers. BRAF interacts with RAS and undergoes a conformational change from an inactive, autoinhibited monomer to an active dimer, which propagates downstream signaling. Because of BRAF’s complex regulation mechanism, the exact order and magnitude of its activation steps have yet to be confirmed experimentally. By studying the inter- and intramolecular interactions of BRAF, we unveil the domain-specific and isoform-specific details of BRAF regulation through pulldown assays, open surface plasmon resonance (OpenSPR), and hydrogen-deuterium exchange mass spectrometry (HDX-MS). We demonstrate that the BRAF specific region (BSR) and cysteine rich domain (CRD) play a crucial role in regulating the activation of BRAF in a RAS isoform-specific manner. Moreover, we quantified the binding affinities between BRAF N-terminal and kinase domains (KD) to reveal their individual roles in autoinhibition. Our findings also indicate that oncogenic BRAF-KD D594G mutant has a lower affinity for the N-terminal domains, implicating that pathogenic BRAF acts through decreased propensity for autoinhibition. Collectively, our study provides valuable insight into the activation mechanism of BRAF kinase to guide the development of new therapeutic strategies for cancer treatment.

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Metadata

Authors
Trebino, T. E., Markusic, B., Nan, H., Banerjee, S., & Wang, Z.
Affiliations
  • Rowan University (US)
  • Max Planck Institute of Biophysics (DE)
  • Wenzhou Medical University (CN)
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2023-12-27
Volume
12
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.88836.3
OpenAlex
W4390266212
Cited by
10
References
4
Citations by year
2026: 3 · 2025: 4 · 2024: 3
FWCI
0.89
Type
journal article
Language
en
Open access
Licence
Locations
eLife (publishedVersion, cc-by)
Funders
  • WW Smith Charitable Trust
  • National Institute of General Medical Sciences (R15GM128099, R01GM138671)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
Keywords
BRAF kinase, Ras isoforms, MAPK signaling, cysteine-rich domain, kinase domain, N-terminal domain, protein-protein interaction

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