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BRAF complexes - Discrete cytosolic macromolecular BRAF complexes exhibit distinct activities and composition

ABSTRACT: BRAF interactomes dependent on the mutation V600E are analyzed by SEC-PCP-SILAC and HA-IPs comparing V600E to WT

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

DISEASE(S): Colon Cancer

SUBMITTER: Joern Dengjel

LAB HEAD: Joern Dengjel

PROVIDER: PXD004585 | Pride | 2017-01-03

REPOSITORIES: Pride

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			Action	DRS
	20120724_BD_48Dox_fw_A01.RAW	Raw
	20120724_BD_48Dox_fw_A02.RAW	Raw
	20120724_BD_48Dox_fw_A03.RAW	Raw
	20120724_BD_48Dox_fw_A04.RAW	Raw
	20120724_BD_48Dox_fw_A05.RAW	Raw

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Discrete cytosolic macromolecular BRAF complexes exhibit distinct activities and composition.

Diedrich Britta B Rigbolt Kristoffer Tg KT Röring Michael M Herr Ricarda R Kaeser-Pebernard Stephanie S Gretzmeier Christine C Murphy Robert F RF Brummer Tilman T Dengjel Jörn J

The EMBO journal 20170116 5

As a central element within the RAS/ERK pathway, the serine/threonine kinase BRAF plays a key role in development and homeostasis and represents the most frequently mutated kinase in tumors. Consequently, it has emerged as an important therapeutic target in various malignancies. Nevertheless, the BRAF activation cycle still raises many mechanistic questions as illustrated by the paradoxical action and side effects of RAF inhibitors. By applying SEC-PCP-SILAC, we analyzed protein-protein interact ...[more]

PMID: 28093501

Similar Datasets

Discrete cytosolic macromolecular BRAF complexes exhibit distinct activities and composition.

Project description:As a central element within the RAS/ERK pathway, the serine/threonine kinase BRAF plays a key role in development and homeostasis and represents the most frequently mutated kinase in tumors. Consequently, it has emerged as an important therapeutic target in various malignancies. Nevertheless, the BRAF activation cycle still raises many mechanistic questions as illustrated by the paradoxical action and side effects of RAF inhibitors. By applying SEC-PCP-SILAC, we analyzed protein-protein interactions of hyperactive BRAFV600E and wild-type BRAF (BRAFWT). We identified two macromolecular, cytosolic BRAF complexes of distinct molecular composition and phosphorylation status. Hyperactive BRAFV600E resides in large complexes of higher molecular mass and activity, while BRAFWT is confined to smaller, slightly less active complexes. However, expression of oncogenic K-RasG12V, either by itself or in combination with RAF dimer promoting inhibitors, induces the incorporation of BRAFWT into large, active complexes, whereas pharmacological inhibition of BRAFV600E has the opposite effect. Thus, the quaternary structure of BRAF complexes is shaped by its activation status, the conformation of its kinase domain, and clinically relevant inhibitors.

| S-EPMC5331759 | biostudies-literature

Transcriptional Response to transgenic BRAF(V600E) in the intestinal epithelium

Project description:BRAF(V600E) is a frequent mutation in colon cancer. We have analysed transcriptional effects of BRAF(V600E) in the intestinal epithelium of transgenic mice that harbour an inducible BRAF(V600E) transgene. Mice used in this experiment were compound transgenic for a stem-cell specific Lgr5-EGFP Reporter and either an inducible TdTomato-2A-BRAF(V600E) transgene or an inducibe TdTomato-luciferase transgene. Transgenes were induced by doxycycline (4mg/ml) treatment provided in a 1% sucrose solution in the drinking water of transgenic mice for 24h. Intestinal crypts were isolated by filtering (70um) following a PBS/EDTA incubation step. Single cell suspensions were made by digestion of crypts in 500ug/ml trypsine and 0.8u/ml DNAseI for approx. 20min at RT. Cell populations for profiling were isolated by FACS, using an Aria SOPR (BD), equipped with a 70um nozzle.

2013-12-31 | E-GEOD-50678 | biostudies-arrayexpress

BRAF interactome - Phospho-proteomic analyses of B-Raf protein complexes reveal new regulatory principles

Project description:SILAC-based analysis of BRAF interaction partners in DT40 chicken cells and mouse embryonic fibroblasts (MEF)

2016-07-13 | PXD003256 | Pride

Oncogenic BRAF regulates oxidative metabolism via PGC1alpha and MITF

Project description:We evaluated two matched human cell lines derived from primary melanocytes, termed pmel+BRAF(V600E) and pmel+BRAF(V600E)+MITF. These cells are therefore isogenic with the exception of the expression of MITF. Gene Set Enrichment Analysis of microarray data identified a highly significant induction of oxidative phosphorylation gene set in MITF expressing cells compared to control cells.

2013-03-07 | E-GEOD-38007 | biostudies-arrayexpress

RNA-seq of BRAF-V600E_mutated and BRAF-V600E_corrected colon cancer organoids

Project description:We aimed to investigate transcriptional changes in human colon cancer organoids with the BRAF-V600E mutation and in human colon cancer organoids in which the BRAF-V600E mutation was corrected by means of CRISPR genome editing. RNAseq was performed at USEQ at the UMC Utrecht (The Netherlands).

2024-02-21 | E-MTAB-13806 | biostudies-arrayexpress

RNA-seq of BRAF-V600E_mutated and BRAF-V600E_corrected colon cancer organoids_advanced starvation

2024-08-23 | E-MTAB-14298 | biostudies-arrayexpress

Illumina Infinium HumanMethylation EPIC beadchip profiling of BRAF-V600E_mutated and BRAF-V600E_corrected colon cancer organoids

Project description:We aimed to investigate genome wide DNA methylation changes in human colon cancer organoids with the BRAF-V600E mutation and in human colon cancer organoids in which the BRAF-V600E mutation was corrected by means of CRISPR genome editing. DNA methylation analysis was performed using the Infinium Methylation EPIC beadchip at the Pathology department of the UMC Utrecht (The Netherlands).

2023-12-20 | E-MTAB-13283 | biostudies-arrayexpress

Anti-BRAF mutation drug resistance enhances EGFR expression in melanomas [expression profiling]

Project description:We treated melanoma cells with BRAF mutation with BRAF inhibitor and screened for BRAF inhibitor resistant cells. We extracted total mRNA from parental cells and resistant cell lines. We compared their expression by carried out Affymetrix Huex 1.0 ST expression array. Two paired parental/derived cell lines were screened as per the summary above. BRAF inhibitory resistant cells had gene expression compared to the parental cell lines.

2014-12-17 | E-GEOD-45648 | biostudies-arrayexpress

RNA-seq of BRAF-wildtype and BRAF-V600E_knock-in healthy colon organoids

Project description:We aimed to investigate transcriptional changes in human healthy colon organoids where the BRAF-V600E mutation was knocked-in by means of CRISPR genome editing. RNAseq was performed at USEQ at the UMC Utrecht (The Netherlands).

2024-03-08 | E-MTAB-13842 | biostudies-arrayexpress

3' RNAseq of EGFR mutant lung cancer cell line overexpressing BRAF V600E treated with inhibitors

Project description:In the human EGFR mutant adenocarcinoma cell line PC9 pBabe empty vector (EV) or BRAF V600E were overexpressed. RNA was extracted from EV, BRAF V600E overexpressing or osimertinib-resistant BRAF V600E expressing cells after 48h treatment with osimertinib, trametinib, a combination of both or vehicle controls. RNA was subjected to 3' UTR RNA sequencing.

2021-10-20 | E-MTAB-11004 | biostudies-arrayexpress

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