Related Experiment Video

Updated: Mar 18, 2026

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
07:49

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods

Published on: July 17, 2019

6.6K

Autophosphorylation on S614 inhibits the activity and the transforming potential of BRAF

Layal Dernayka1, Nora Rauch2, Mohamed-Ali Jarboui1

  • 1Medical Proteome Center, Division for Experimental Ophthalmology, Institute for Ophthalmic Research, University of Tuebingen, Tuebingen, Germany.

Cellular Signalling
|June 28, 2016
PubMed

Insights

Researchers compared BRAF and BRAF V600E phosphorylation patterns using mass spectrometry. They found S614 phosphorylation is enhanced in BRAF V600E, suggesting an inhibitory role in BRAF signaling.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Signal Transduction

Background:

  • BRAF (proto-oncogene serine/threonine-protein kinase) is a key regulator of the MAPK/ERK pathway.
  • BRAF mutations, particularly V600E, are prevalent in approximately 8% of human cancers.
  • The MAPK/ERK pathway controls fundamental cellular processes including growth, survival, differentiation, and transformation.

Purpose of the Study:

  • To quantitatively map and compare phosphorylation site patterns between wild-type BRAF and the BRAF V600E mutant.
  • To investigate the functional consequences of differential phosphorylation, specifically at S614.

Main Methods:

  • Quantitative mass spectrometry was employed to analyze and compare global phosphorylation profiles.
  • Site-directed mutagenesis was used to assess the impact of S614 phosphorylation.

Main Results:

  • Identified shared and differentially phosphorylated sites between BRAF and BRAF V600E.
  • Observed a significant ~5-fold increase in S614 phosphorylation in BRAF V600E compared to wild-type BRAF.
  • Demonstrated that S614 phosphorylation is mitogen-inducible, results from autophosphorylation, and its mutation enhances kinase activity and transforming potential.

Conclusions:

  • Phosphorylation at S614 acts as an inhibitory mechanism, contributing to the physiological regulation and shut-down of BRAF signaling.
  • Understanding these phosphorylation dynamics is crucial for comprehending BRAF-driven oncogenesis and developing targeted therapies.
Keywords:
BRAFInhibition of kinase activity and transformationS614 autophosphorylation

More Related Videos

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells06:44

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

Published on: March 1, 2024

1.9K
Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation15:05

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Published on: May 20, 2020

9.4K

Related Concept Videos

MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
9.1K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
13.7K
The Ras Gene02:38

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...
7.5K
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
6.2K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
19.0K
mTOR Signaling and Cancer Progression03:03

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
5.0K