Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Interactome dynamics of RAF1-BRAF kinase monomers and dimers

ABSTRACT: RAF kinases play major roles in cancer. BRAFV600E mutants drive ~6% of human cancers. Potent kinase inhibitors exist but show variable effects in different cancer types, sometimes even inducing paradoxical RAF kinase activation. Both paradoxical activation and drug resistance are frequently due to enhanced dimerization between RAF1 and BRAF, which maintains or restores the activity of the downstream MEK-ERK pathway. Here, using quantitative proteomics we mapped the interactomes of RAF1 monomers, RAF1-BRAF and RAF1-BRAFV600E dimers identifying and quantifying >1,000 proteins. In addition, we examined the effects of vemurafenib and sorafenib, two different types of clinically used RAF inhibitors. Using regression analysis to compare different conditions we found a large overlapping core interactome but also distinct condition specific differences. Given that RAF proteins have kinase independent functions such dynamic interactome changes could contribute to their functional diversification. Analysing this dataset may provide a deeper understanding of RAF signalling and mechanisms of resistance to RAF inhibitors.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Kidney Epithelium, Hek-293 Cell

SUBMITTER: Kieran Wynne

LAB HEAD: Walter Kolch

PROVIDER: PXD036792 | Pride | 2023-02-27

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Raf/MEK signatures in zebrafish liver cells

Project description:An inducible oncogenic cell line ZFL-?Raf1-ER was established, in which oncogenic human Raf-1(?Raf1) can be activated in zebrafish liver cells by administration of 4-hydroxytamoxifen (4HT). Transcriptional profilling of the ZFL-?Raf1-ER cells with and without administration of 4HT and/or the MEK inhibitor U0126 defined the gene signatures transcriptionally regulated by hyperactive Raf/MEK/MAPK signaling in zebrafish liver cells. The ZFL-?Raf1-ER cells were cultured in 12-well plates till confluence. Cell were starved for 4h in plain medium and susequently treated with or without 1µM 4HT and/or 30µM U0126 for 12h. Biological triplications were taken for each condition.

2011-07-07 | E-GEOD-29308 | biostudies-arrayexpress

Time-course expression data from HEK293âRAF1:ER cells stimulated with 4OHT, U0126, CYHX, ActD, EGF, FGF, or IGF

Project description:We integrate experimental data and mathematical modelling to unveil how ERK signal duration is relayed to mRNA dynamics. HEK293 cells were transfected with an inducible form of RAF (âRAF1:ER) and induced with tamoxifen (4OHT) or stimulated with different growth factors (EGF, FGF, iGF). Effects of RAF signalling were perturbed subsequently or in parallel with MEK inhibtor U0126, translation inhibtor cycloheximide (CYHX) and actinomycin D (ActD).

2017-04-28 | E-GEOD-72919 | biostudies-arrayexpress

Gene expression data from cultured cells or from nude mice xenografts

Project description:In this study we performed gene transcription analysis of parental MACF7 cell and MCF7 engineered to overexpress oncoprotein RAF1. We demonstrate that overexpression of RAF1 leads to development of ivasive phenotype. Total RNA was extracted from culturued cells or from nude mice xenografts.

2011-10-14 | E-GEOD-29884 | biostudies-arrayexpress

RNA-seq of canine MDCK epithelial cell line compared to cell line overexpressing ERK2 or cell line with conditional activation of the ERK pathway

Project description:Using RNA-seq and canine MDCK epithelial cells, we analyzed mRNA expression profiles of cells overexpressing ERK2 and cells where the ERK pathway was activated for 24 hrs. For this purpose, three cell lines were used: parental MDCK cells, MDCK cells overexpressing FLAG-ERK2 or MDCK cells expressing conditionally active Raf protein (ΔRaf1:ER). Activation of the ERK pathways was achieved by the addition of 4-Hydroxytamoxifen that converts ΔRaf1:ER protein to the active form and it subsequently activates the ERK pathway.

2022-10-01 | E-MTAB-12132 | biostudies-arrayexpress

CXMS BS3 Ras/BRAF/MEK/1433 complex

Project description:RAF-family kinases are activated by recruitment to the plasma membrane by GTP-bound RAS, where they initiate signaling through the MAP kinase cascade. Here we crosslinked (BS3), digested (trypsin), and analyzed via LC-MS/MS KRAS (human, residues 1-169) bound to intact BRAF (human) in an autoinhibited state with MEK1 (human, ) and a 14-3-3 (SF9) dimer . Analysis of this KRAS/BRAF/MEK1/14-3-3 complex reveals both interprotein and intraprotein crosslinks.

2023-10-24 | PXD042584 | Pride

iMixPro: intelligent Mixing of Proteomes for elimination of false positives in affinity purification-mass spectrometry

Project description:Protein complexes are essential in most organizational and functional aspects of the cell. Different strategies currently exist to analyze such protein complexes by mass spectrometry, including affinity purification (AP-MS) and proximal labeling based strategies. However, the high sensitivity of current mass spectrometers typically results in extensive protein lists mainly consisting out of non-specifically co-purified proteins. Finding the true positive interactors in these lists thus remains highly challenging with proposed solutions on all levels of such experiments. Here, we report an intelligent experimental design based on differential isotopic labeling combined with non-equal mixing of control and experimental samples to discover bona fide interaction partners in AP-MS experiments. We apply this intelligent Mixing of Proteomes (iMixPro) concept to overexpression experiments where we investigate the interactomes of RAF1, RNF41 and TANK, and also to engineered cell lines expressing epitope-tagged endogenous PTPN14, JIP3 and IQGAP1. For all baits, we observed both known and novel interactions. The results for RNF41 and TANK were compared to a classical affinity purification design which clearly underscores the efficiency and the specificity of the iMixPro approach.

2016-09-27 | PXD004246 | Pride

Cardiac transcriptome analysis of RKIP-transgenic and GRK2-transgenic mice by NGS

Project description:The RAF kinase inhibitor protein, RKIP, is a dual inhibitor of the RAF1 kinase and the G-protein-coupled receptor kinase 2 (GRK2). By inhibition of the proto-oncogenic and pro-survival RAF1-MAPK pathway, the RAF kinase inhibitor protein, RKIP, acts as a tumor suppressor, which enhances cardiomyocyte death and promotes the development of symptoms of heart failure. To elucidate pathomechanisms of heart failure induced by RKIP, the study determined the cardiac transcriptomes of eight-month-old, male, transgenic mice with cardiac-specific expression of RKIP (PEBP1) under control of the myocardium-specific, alpha-MHC promoter. In addition, the study determined the cardiac transcriptomes of GRK2-transgenic mice. Tg-GRK2 mice have a slightly increased transgenic expression of GRK2. According to NGS data, cardiac GRK2-Grk2 transcript levels of Tg-GRK2 mice are 1.59±0.10-fold higher than those of non-transgenic FVB hearts. In Tg-GRK2 mice, transgenic GRK2 is expressed under control of the ubiquitous CMV immediate-early promoter/enhancer. The non-transgenic control group are age-matched, male, nontransgenic FVB/N mice. NGS data of this study document transcriptome changes underlying the heart failure phenotype induced by transgenic RKIP expression and cardiac degeneration induced by GRK2 expression.

2021-12-27 | GSE191316 | GEO

Gene expression profiling of colorectal cancers to understand molecular changes and clinicopathological features

Project description:Gene expression was analyzed in terms of canonical molecular changes and clinicopathological features to elucidate alternative or subordinate pathways during colorectal tumorigenesis and tumor growth. Eighty-four sporadic colorectal cancer patients, standardized by tumor location, were consecutively enrolled. Representative molecular changes including APC, TP53, Wnt, RAF, and mismatch repair defect (MMR) were recorded for each sample. Keywords: disease state analysis; sub-type analysis within colorectal cancers 84 samples from colorectal patients were analyzed. Paired tumor and adjacent normal tissues from the same patient were used for hybridization onto custom-made, 21k dual channel cDNA arrays. We prepared a similar number of samples from each of the three tumor locations (ascending 27, descending 29, and rectum 28) and recorded for each sample important molecular changes such as APC, TP53, RAF, WNT, and MMR mutations.

2009-02-28 | E-GEOD-10982 | biostudies-arrayexpress

Endothelial Raf1 activation regulates arterial morphogenesis

Project description:Analyze effect of Raf1 S259A on gene expression in HUVEC Total RNA obtained from HUVEC infected with empty lentiviral vector pLVX-IRES-puro (control) or that expressing human RAF1 WT (WT) or RAF1 S259A (S259A).

2012-12-18 | E-GEOD-42964 | biostudies-arrayexpress

Mass spectrometry analysis following chromatin fractionation in growing and senescent cells

Project description:This project contains the raw and result files for a mass spectrometry analysis comparing the abundance of proteins following chromatin fractionation in growing cells and in senescent cells for the WI-38hTERT/GFP-RAF1-ER cell line. Two replicas were analysed for each cell condition. The data are used to support a study on using polymer modelling to investigate the roles of heterochromatin and lamina interactions on chromosome reorganisation in senescence.

2019-09-25 | PXD014929 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data