Related Experiment Video

Updated: Jul 20, 2026

Bioluminescence Resonance Energy Transfer (BRET)-Based Assay for Measuring Interactions of CRAF with 14-3-3 Proteins in Live Cells
06:44

Bioluminescence Resonance Energy Transfer (BRET)-Based Assay for Measuring Interactions of CRAF with 14-3-3 Proteins in Live Cells

Published on: March 1, 2024

Regulation of ERK3/MAPK6 expression by BRAF

Klaus P Hoeflich1, Michael T Eby, William F Forrest

  • 1Department of Molecular Biology, Genentech, South San Francisco, CA 94080, USA.

International Journal of Oncology
|September 12, 2006
PubMed

Insights

Activating BRAF mutations drive cancer. This study identifies novel genes regulated by BRAF signaling, including extracellular signal-regulated kinase-3 (ERK3), a potential marker for BRAF-targeted therapies.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Activating mutations in the serine/threonine kinase BRAF, particularly the BRAF V600E mutation, are common in various cancers.
  • These mutations lead to increased kinase activity, aberrant downstream signaling, and cellular transformation, driving oncogenesis.

Purpose of the Study:

  • To comprehensively analyze the transcriptional program regulated by oncogenic BRAF signaling.
  • To identify novel genes involved in BRAF-mediated oncogenesis.
  • To investigate the role of extracellular signal-regulated kinase-3 (ERK3/MAPK6) in BRAF-driven signaling.

Main Methods:

  • Microarray gene expression profiling of cells with conditionally active BRAF V600E.
  • Analysis of novel gene expression changes.
  • Investigation of ERK3 protein stability and regulation.
  • RNA interference (RNAi) mediated knockdown of BRAF and MEK inhibitor treatment in melanoma cells.

Main Results:

  • Identified novel genes influencing proliferation, cell survival, angiogenesis, and immune surveillance as mediators of BRAF oncogenic signaling.
  • Demonstrated high expression of extracellular signal-regulated kinase-3 (ERK3/MAPK6) in response to BRAF signaling.
  • Showed that BRAF inhibition leads to rapid ERK3 degradation.
  • Confirmed that elevated ERK3 expression is mediated through MEK1/2 signaling in melanoma cells.

Conclusions:

  • BRAF signaling regulates a distinct transcriptional program involving novel oncogenic mediators.
  • Extracellular signal-regulated kinase-3 (ERK3) is a novel downstream target of BRAF/MEK signaling.
  • ERK3 represents a potential pharmacodynamic marker for monitoring BRAF-targeted therapies in melanoma.

More Related Videos

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods07:49

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

Published on: July 17, 2019

Assaying Protein Kinase Activity with Radiolabeled ATP08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

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...
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 rapamycin-insensitive companion...
Master Transcription Regulators02:23

Master Transcription Regulators

Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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...
Interactions Between Signaling Pathways01:19

Interactions Between Signaling Pathways

Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
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...