Affinage

ARAF

Serine/threonine-protein kinase A-Raf · UniProt P10398

Length
606 aa
Mass
67.6 kDa
Annotated
2026-06-09
100 papers in source corpus 36 papers cited in narrative 35 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ARAF is a serine/threonine kinase of the RAF family that functions in the RAS–RAF–MEK–ERK cascade but possesses intrinsically lower MEK kinase activity than BRAF or CRAF, phosphorylating MEK1 hundreds-fold less efficiently in vitro (PMID:7565795). This low basal activity is set by isoform-specific N-region constraints, where residues such as Y296 and G300 restrain catalytic output and engineered substitutions can render the kinase constitutively active (PMID:17613527), with additional positive regulation supplied by an isoform-specific hinge segment and a MEK-binding serine (S432) identified by phosphosite mapping (PMID:18662992). ARAF selectively activates MEK1 but not MEK2 downstream of growth factor receptors (PMID:8621729), and its activation is dimerization-dependent: it homo- and heterodimerizes, and dimer-deficient mutants fail to bind MEK1 or activate ERK (PMID:25097033). Genetic loss-of-function studies show ARAF is largely dispensable alone—knockout MEFs retain normal ERK signaling with compensatory BRAF/CRAF upregulation (PMID:11821947)—but ARAF and RAF-1 act redundantly to drive transient MEK/ERK phosphorylation and G1/S progression (PMID:15856007), and ARAF scaffolds and stabilizes BRAF:CRAF heterodimers (PMID:22926515). Beyond canonical kinase signaling, ARAF carries out several kinase-independent functions: it sequesters and inactivates the pro-apoptotic kinase MST2 to block apoptosis, a function gated by mitochondrial versus plasma-membrane localization controlled by KSR2 (PMID:20145135, PMID:26891695); it activates RAS by binding RAS and displacing the GAP NF1 to raise RAS-GTP (PMID:35613620); and it suppresses ERBB3 transcription via KLF5 (PMID:35302851). ARAF also directly phosphorylates Smad2 in its linker region (S253) in a MEK/ERK-independent manner to attenuate Nodal/Smad2 signaling during development (PMID:23591895). Alternative splicing produces a truncated dominant-negative isoform that retains the RAS-binding domain and suppresses RAS-ERK signaling, regulated by hnRNP H, hnRNP A2 and c-Myc (PMID:21512137, PMID:24572810). Recurrent gain-of-function kinase-domain mutations (e.g. S214P/S214C, N217I) drive oncogenic MAPK activation and a lymphatic/lymphangiogenic phenotype rescued by MEK inhibition (PMID:24569458, PMID:31263281), and ARAF kinase-domain mutations confer resistance to type II RAF dimer inhibitors in a manner requiring both kinase activity and dimerization (PMID:33953400, PMID:33355204).

Mechanistic history

Synthesis pass · year-by-year structured walk · 21 steps
  1. 1995 High

    Established that ARAF is a functional RAF kinase but quantitatively the weakest activator of MEK, defining the central puzzle of its distinct regulation.

    Evidence Inducible Raf:ER fusion system with direct in vitro kinase comparison across all three RAF isoforms

    PMID:7565795

    Open questions at the time
    • Did not explain the structural basis of low activity
    • Did not identify ARAF-specific substrates beyond MEK
  2. 1996 Medium

    Defined ARAF as a MEK1-selective kinase coupled to specific receptor and second-messenger inputs distinct from c-Raf, showing isoform specialization at the substrate and regulatory level.

    Evidence Yeast two-hybrid, in vitro kinase assays, and pharmacological perturbation in HeLa and cardiomyocytes

    PMID:7592840 PMID:8621729

    Open questions at the time
    • MEK1 vs MEK2 selectivity mechanism not structurally resolved
    • Single-lab biochemical readouts
  3. 1997 High

    Identified the kinase-domain regulatory residues and CK2β as activity modulators, beginning to explain how ARAF catalytic output is constrained and tuned.

    Evidence Site-directed mutagenesis with transformation/kinase readouts; yeast two-hybrid and Sf9 co-expression kinase assays

    PMID:9042965 PMID:9042966 PMID:9285556

    Open questions at the time
    • In vivo relevance of CK2β regulation not established
    • Physiological phosphorylation state of the regulatory residues unmapped
  4. 1999 Medium

    Placed ARAF as the primary Raf coupling PI3-K to MEK-ERK in hematopoietic cells, distinguishing its upstream wiring from BRAF/CRAF.

    Evidence PI3-K inhibitors, dominant-negative isoforms, and IP kinase assays in IL-3-dependent cells

    PMID:10066754

    Open questions at the time
    • Molecular basis of PI3-K-to-ARAF coupling not defined here
    • Cell-type generality unknown
  5. 2000 Medium

    Revealed ARAF subcellular targeting (mitochondria) and a phosphotyrosine-independent association with PI3-kinase p85, hinting at non-canonical localization and binding modes.

    Evidence Mitochondrial fractionation with immunogold EM; phage display, co-IP and in vitro binding for p85

    PMID:10848612 PMID:10967104

    Open questions at the time
    • Functional consequence of mitochondrial localization unresolved at this stage
    • hTOM/hTIM interactions not functionally validated
  6. 2002 High

    Showed ARAF is dispensable for ERK signaling alone due to BRAF/CRAF compensation, reframing its role as specialized or redundant rather than essential to the core cascade.

    Evidence Gene-targeted ARAF knockout MEFs and ES cells with functional readouts

    PMID:11821947

    Open questions at the time
    • Did not test combined RAF loss
    • Tissue-specific or non-ERK roles not addressed
  7. 2003 Medium

    Identified the first kinase-independent anti-apoptotic role of ARAF via MST2 sequestration and an ARAF-specific inhibitor TH1, opening non-canonical signaling functions.

    Evidence siRNA knockdown with MST2 epistasis and apoptosis assays; co-IP and kinase assays for TH1 binding

    PMID:14684750 PMID:20145135

    Open questions at the time
    • Structural basis of MST2 sequestration unresolved
    • Single-lab findings
  8. 2005 High

    Demonstrated functional redundancy of ARAF with RAF-1 in driving transient ERK signaling and cell cycle entry, resolving the discrepancy with the single-knockout phenotype.

    Evidence ARAF/RAF-1 double-knockout MEFs with cell cycle and phospho-MEK/ERK analysis; lipid binding assays

    PMID:15736953 PMID:15856007

    Open questions at the time
    • Mechanism of selectivity for transient vs sustained ERK unclear
    • Physiological role of phosphoinositide binding not established
  9. 2007 Medium

    Mapped the N-region residues that impose ARAF's low basal activity, providing a structural rationale for its weak kinase output relative to other RAFs.

    Evidence Site-directed mutagenesis with kinase assays and modeling from BRAF structure; co-IP metabolic studies for M2-PK

    PMID:17613527 PMID:18225557

    Open questions at the time
    • No crystal structure of ARAF kinase domain
    • M2-PK interaction is low-confidence and mechanistically thin
  10. 2008 Medium

    Defined in vivo ARAF phosphosites controlling MEK binding (S432) and activation (IH-segment), refining the activation logic of the kinase.

    Evidence Mass spectrometry plus mutagenesis with in vitro kinase assays and EGF stimulation

    PMID:18662992

    Open questions at the time
    • Kinases responsible for these phosphorylations not identified
    • Single-lab phosphosite mapping
  11. 2011 Medium

    Established that c-Myc/hnRNP H-controlled splicing produces a dominant-negative ARAF(short) that suppresses RAS-ERK, linking isoform choice to pathway output.

    Evidence c-Myc and hnRNP H manipulation, splice-isoform RT-PCR, and RAS-GTP/ERK assays

    PMID:21512137

    Open questions at the time
    • Physiological triggers of splicing switch in vivo unclear
    • Quantitative isoform ratios in tissues not defined
  12. 2012 Medium

    Showed ARAF is not redundant with CRAF and scaffolds BRAF:CRAF heterodimers under RAF inhibitor conditions, defining a structural role distinct from catalysis.

    Evidence Co-IP, knockdown/overexpression, RAF inhibitor treatment, and transformation assays

    PMID:22926515

    Open questions at the time
    • Dimer interface stoichiometry not fully resolved
    • Endogenous relevance vs overexpression unclear
  13. 2013 High

    Identified Smad2 as a direct MEK-independent ARAF substrate, establishing a developmental signaling role outside the canonical MAPK cascade.

    Evidence In vitro kinase assay with S253 mutagenesis and zebrafish knockdown with rescue

    PMID:23591895

    Open questions at the time
    • Regulation of ARAF-Smad2 activity in mammalian development not mapped
    • Crosstalk with MEK-ERK substrate selection unresolved
  14. 2014 High

    Demonstrated that ARAF activation strictly requires dimerization, unifying its inhibitor-induced and growth-factor-driven activation with a shared mechanistic requirement.

    Evidence Recombinant protein competition, dimer-deficient mutants, siRNA, and 3D spheroid invasion assays

    PMID:25097033

    Open questions at the time
    • Identity of obligate dimer partners in vivo not fully defined
    • Dimer-specific conformational change not structurally captured
  15. 2014 Medium

    Identified recurrent oncogenic ARAF kinase-domain mutations in LCH, lung, and other cancers that produce hyperactive, drug-targetable MAPK kinases.

    Evidence In vitro kinase and transformation assays with vemurafenib/sorafenib sensitivity; whole-genome sequencing

    PMID:24569458 PMID:24652991

    Open questions at the time
    • Mutation-specific mechanism of activation not fully resolved
    • Clinical response durability not addressed
  16. 2014 Medium

    Showed hnRNP A2-driven splicing reduces dominant-negative ARAF and activates RAS-MAPK in hepatocellular carcinoma, reinforcing splicing as an oncogenic switch.

    Evidence hnRNP A2 siRNA, splice-isoform RT-PCR, EGF-induced ERK assays, and tumor formation

    PMID:24572810

    Open questions at the time
    • Generality across tumor types not tested
    • Direct hnRNP A2 binding sites on Araf pre-mRNA not mapped
  17. 2015 Medium

    Confirmed additional oncogenic ARAF mutations (N217I, G322S) drive MAPK activation in cholangiocarcinoma, extending the mutational spectrum.

    Evidence RNA/exome sequencing with MAPK activation and transformation assays

    PMID:25608663

    Open questions at the time
    • Differential potency of mutations not benchmarked
    • Therapeutic implications not tested in vivo
  18. 2016 Medium

    Showed localization (mitochondria vs plasma membrane, set by KSR2) determines whether ARAF blocks or permits MST2-mediated apoptosis, linking spatial control to cell fate.

    Evidence Confocal imaging, fractionation, KSR2 knockdown, and apoptosis assays in a differentiation model

    PMID:26891695

    Open questions at the time
    • Mechanism of KSR2-controlled relocalization not defined
    • In vivo relevance during tissue differentiation untested
  19. 2019 High

    Defined the recurrent S214P gain-of-function mutation as a cause of lymphatic anomalies driven by ERK hyperactivation and reversible by MEK inhibition, establishing a disease-relevant mechanism.

    Evidence Lentiviral expression, ERK/lymphangiogenesis/junction assays, zebrafish model, and trametinib rescue

    PMID:31263281

    Open questions at the time
    • Why S214 loss is specifically activating not structurally resolved
    • Endothelial-specific cofactors not identified
  20. 2020 High

    Established ARAF as a clinically important driver of resistance to type II RAF dimer inhibitors, requiring both kinase activity and dimerization, and showing ARAF can paradoxically maintain MAPK signaling under inhibition.

    Evidence Resistant cell lines, patient ctDNA sequencing, dimerization/kinase mutants, and xenografts across multiple RAF inhibitors

    PMID:33355204 PMID:33953400

    Open questions at the time
    • Strategies to co-target ARAF dimers not validated clinically
    • Why ARAF is spared by these inhibitors structurally not fully defined
  21. 2022 High

    Uncovered kinase-independent ARAF functions—RAS activation via NF1 displacement and ERBB3 suppression via KLF5—that broaden its role beyond a MEK kinase and explain inhibitor and EGFR-therapy resistance.

    Evidence Co-IP of ARAF-RAS-NF1 complexes, RAS-GTP pull-downs, kinase-dead mutants, promoter reporters, and rescue experiments

    PMID:35139374 PMID:35302851 PMID:35613620

    Open questions at the time
    • Structural basis of NF1 displacement not solved
    • Balance between ARAF's kinase and scaffold functions in tumors unclear

Open questions

Synthesis pass · forward-looking unresolved questions
  • How ARAF's multiple kinase-dependent and kinase-independent activities are integrated and spatially partitioned within a single cell, and whether its non-canonical functions are therapeutically separable, remains unresolved.
  • No high-resolution structure of full-length ARAF or its dimers
  • Unknown how localization, splicing, and partner choice are co-regulated
  • Relative contribution of kinase vs scaffold roles in disease undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016740 transferase activity 3 GO:0098772 molecular function regulator activity 3 GO:0140096 catalytic activity, acting on a protein 3 GO:0008289 lipid binding 1
Localization
GO:0005739 mitochondrion 2 GO:0005886 plasma membrane 2 GO:0005768 endosome 1
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-1643685 Disease 3 R-HSA-5357801 Programmed Cell Death 2 R-HSA-1640170 Cell Cycle 1
Partners
BRAFCRAFKSR2MEK1MST2NF1RASSMAD2
Complex memberships
ARAF-RAS-NF1 complexBRAF:CRAF heterodimer

Evidence

Reading pass · 35 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1995 A-Raf kinase domain (deltaA-Raf:ER) activates MEK and ERK upon estradiol-induced activation in mammalian cells, but phosphorylates MEK1 in vitro at least 500-fold less efficiently than deltaB-Raf:ER and ~10-fold less efficiently than deltaRaf-1:ER, demonstrating A-Raf has intrinsically lower MEK kinase activity than the other Raf isoforms. Inducible Raf:ER fusion protein system; immune-complex in vitro kinase assays; Western blot for MEK and ERK activation Molecular and cellular biology High 7565795
1996 A-Raf (and c-Raf) expressed in neonatal rat ventricular myocytes both phosphorylate, activate, and form complexes with MEK1 in vitro; however, A-Raf and c-Raf are differentially regulated by hypertrophic stimuli: TPA produces sustained A-Raf activation, whereas aFGF activates c-Raf but not A-Raf, and cAMP-dependent PKA inhibits c-Raf but not A-Raf. Immunoprecipitation kinase assays; co-complex formation; pharmacological activation/inhibition in cultured cardiomyocytes The Journal of biological chemistry Medium 7592840
1996 A-Raf selectively phosphorylates and activates MEK1 but not MEK2 in response to EGF stimulation in HeLa cells; yeast two-hybrid using MEK1 S218/222A mutant as bait identified all three Raf family members as interactors, suggesting elimination of phosphorylation sites stabilizes the kinase–substrate interaction. Yeast two-hybrid; in vitro kinase assay; EGF stimulation of HeLa cells; MEK1/MEK2 phosphorylation assays The Journal of biological chemistry Medium 8621729
1997 The regulatory subunit of protein kinase CK2 (CK2β) specifically binds to A-Raf (residues 550–569 in the kinase domain) but not to B-Raf or C-Raf, and co-expression of CK2β with A-Raf in Sf9 cells enhances A-Raf kinase activity ~10-fold; this effect is abolished by the CK2 catalytic α subunit. Yeast two-hybrid screening; Sf9 co-expression kinase assays; domain mapping FEBS letters Medium 9042965 9042966
1997 Mutation of two key tyrosine residues (analogous to activating Asp residues in B-Raf) to aspartic acid in A-Raf's kinase domain (~10-fold increase in catalytic activity) or to phenylalanine (loss of activity) demonstrates that these tyrosines are critical regulatory residues controlling A-Raf catalytic activity; introduction of Asp increases oncogenic transformation, while Phe abolishes it. Site-directed mutagenesis; Raf:ER fusion protein transformation assays in Rat1/3T3 cells; immune-complex kinase assays; Sf9 insect cell expression Oncogene High 9285556
1999 In IL-3-dependent hematopoietic cells, A-Raf (but not B-Raf or c-Raf) is activated downstream of PI3-K, is insensitive to cAMP inhibition, and is required for MEK-ERK activation; dominant-negative A-Raf blocks MEK activation whereas dominant-negative c-Raf does not, establishing A-Raf as the primary Raf isoform coupling PI3-K signals to MEK in these cells. PI3-K inhibitor (wortmannin/LY294002); cAMP elevation; dominant-negative isoform overexpression; immunoprecipitation kinase assays The Journal of biological chemistry Medium 10066754
2000 A-RAF localizes specifically to mitochondria in rat liver cells (determined by Western blot and immunogold labeling of purified mitochondria), and two novel human mitochondrial import receptor proteins (hTOM and hTIM, related to TOM/TIM translocase components) interact specifically with A-RAF; 14-3-3, which interacts with C-RAF, does not bind A-RAF's N-terminal domain. Western blot of purified mitochondrial fractions; immunogold electron microscopy; yeast two-hybrid; protein interaction studies Molecular and cellular biology Medium 10848612
2000 A-Raf associates with the p85 subunit of PI3-kinase through a phosphotyrosine-independent SH2 domain interaction requiring basic residues (part of the sequence LQRIRS) in A-Raf; this complex exists in both quiescent and growth factor-stimulated cells and can be reconstituted by direct in vitro binding. Phage display peptide library; co-immunoprecipitation; in vitro direct binding assay; mutagenesis of basic residues The Journal of biological chemistry Medium 10967104
2002 A-Raf-deficient mouse embryonic fibroblasts and ES cells show no defects in ERK activation, proliferation, differentiation, apoptosis, or transformation by oncogenic Ras or Src; B-Raf and Raf-1 activities toward MEK are both significantly increased in A-Raf-deficient MEFs, suggesting compensatory upregulation. Gene-targeted A-Raf knockout MEFs and ES cells; ERK activation assays; transformation assays; immunoprecipitation kinase assays Oncogene High 11821947
2003 A-Raf prevents cancer cell apoptosis by sequestering and inactivating the pro-apoptotic MST2 kinase; siRNA knockdown of A-Raf results in MST2-dependent apoptosis, and enforced A-Raf expression counteracts etoposide-induced apoptosis; this anti-apoptotic function requires expression of the splicing factor hnRNP H for correct A-Raf transcription. siRNA knockdown; apoptosis assays; enforced expression; epistasis with MST2 Cancer research Medium 20145135
2005 A-Raf and Raf-1 have a combined role in controlling transient (but not sustained) ERK/MEK phosphorylation and G1/S cell cycle progression; double knockout (A-Raf/Raf-1) MEFs show delayed S-phase entry, reduced transient MEK/ERK phosphorylation, and reduced c-Fos and cyclin D1 expression, while single knockouts are unaffected. Double knockout mouse embryonic fibroblasts; cell cycle analysis; Western blot for phospho-MEK/ERK; Ki67 staining Oncogene High 15856007
2005 A-Raf binds to phosphoinositides including PI(3)P, PI(4)P, PI(5)P, PI(3,5)P2, PI(4,5)P2, PI(3,4)P2, and PA; mutagenesis of two basic residues K50 and R52 in the Ras-binding domain abolishes PI(4,5)P2 binding, suggesting these residues are the PI(4,5)P2 binding site; a second phosphoinositide binding region resides between residues 200–606. Lipid binding assays; site-directed mutagenesis (K50, R52); deletion mutant analysis Biochemistry Medium 15736953
2007 A-Raf interacts with and regulates pyruvate kinase M2 (M2-PK): in primary mouse fibroblasts, A-Raf induces M2-PK dimerization (inactive form) reducing glycolysis, whereas oncogenic A-Raf in NIH3T3 cells increases the tetrameric (active) form of M2-PK promoting glycolytic energy production. Co-immunoprecipitation; enzyme activity assays; glycolysis measurements; primary vs. immortalized fibroblasts comparison Anticancer research Low 18225557
2007 The N-region of A-RAF (specifically residues including Y296 and G300) determines its low basal kinase activity and limited inducibility; substitution Y296R in A-RAF yields constitutively active kinase, while G300S (mimicking B-/C-RAF) is inhibitory; conversely, S339G in C-RAF (mimicking A-RAF) creates constitutively active C-RAF, implicating a conserved N-region residue (S339 in C-RAF/R398 interaction) as an inhibitory constraint. Site-directed mutagenesis; kinase activity assays; molecular modeling based on B-RAF crystal structure The Journal of biological chemistry Medium 17613527
2008 Mass spectrometry identified multiple novel in vivo phosphorylation sites in A-RAF; Ser-432 participates in MEK binding and is indispensable for A-RAF signaling; an isoform-specific hinge segment (IH-segment, residues 248–267) containing Ser-257, Ser-262, and Ser-264 positively regulates A-RAF activation; phosphorylation within the activation segment does not contribute to EGF-mediated activation. Mass spectrometry; site-directed mutagenesis; in vitro kinase assays; EGF stimulation The Journal of biological chemistry Medium 18662992
2009 A-RAF kinase function in ARF6-regulated endocytic membrane traffic: an N-terminal A-RAF fragment (AR149, corresponding to splice variant DA-RAF2) co-localizes with ARF6 on tubular endosomes and exerts dominant-negative effects on endocytic recycling (not internalization); A-RAF-mediated ERK activation via MEK is required for ARF6-dependent recycling to the recycling compartment. Co-localization microscopy; dominant-negative expression; endocytic trafficking assays; A-RAF depletion; MEK-ERK pathway inhibition PloS one Medium 19247477
2011 c-Myc positively controls hnRNP H expression, which in turn regulates A-raf mRNA splicing to produce full-length A-Raf vs. A-Raf(short), a truncated dominant-negative isoform retaining the Ras-binding domain; A-Raf(short) suppresses Ras activation and ERK signaling; full-length A-Raf inhibits MST2-dependent apoptosis, while A-Raf(short) does not regulate MST2. c-Myc manipulation; hnRNP H expression analysis; RT-PCR for splice isoforms; dominant-negative overexpression; Ras-GTP and ERK activation assays Cancer research Medium 21512137
2012 ARAF acts as a scaffold to stabilize BRAF:CRAF heterodimers in cells treated with RAF inhibitors; ARAF binds to and is activated by BRAF; ARAF is not functionally redundant with CRAF and cannot substitute for CRAF downstream of RAS; two cancer-associated ARAF mutations inactivate the kinase. Co-immunoprecipitation; ARAF knockdown/overexpression; RAF inhibitor treatment; kinase activity assays; transformation assays Oncogene Medium 22926515
2013 Araf directly phosphorylates Smad2 in the linker region (S253 indispensable) in a MEK/ERK-independent manner, promoting degradation of activated Smad2 and attenuating Nodal/Smad2 signaling; in zebrafish, araf knockdown increases activated Smad2, expands mesendoderm, and causes dorsalization. In vitro kinase assay; site-directed mutagenesis (S253); zebrafish knockdown; Smad2 phosphorylation and degradation assays; co-immunoprecipitation Nature communications High 23591895
2014 ARAF dimerization is required for MAPK pathway activation and cell migration: RAF inhibitors induce ARAF homodimerization; ARAF mutants unable to homodimerize fail to interact with endogenous MEK1 and do not activate MEK1 or ERK1/2; in purified protein solution, all three RAF isoforms compete for binding to MEK1; ARAF knockdown prevents RAF-inhibitor-induced MEK1 and ERK1/2 activation and tumor cell spheroid invasion. Recombinant protein competition assay; co-immunoprecipitation; dimer-deficient ARAF mutant overexpression; siRNA knockdown; 3D spheroid invasion assay Science signaling High 25097033
2014 Compound kinase-domain mutations in ARAF found in LCH create a highly active MAP kinase kinase in vitro and transform mouse embryo fibroblasts; mutant ARAF activity is inhibited by the BRAF inhibitor vemurafenib. In vitro kinase assay; mouse embryo fibroblast transformation assay; vemurafenib inhibition Blood Medium 24652991
2014 ARAF mutations S214C and nearby residues transform immortalized human airway epithelial cells in a sorafenib-sensitive manner, demonstrating these mutations confer oncogenic gain-of-function kinase activity. Cell transformation assay; sorafenib sensitivity assay; whole-genome sequencing The Journal of clinical investigation Medium 24569458
2014 hnRNP A2 regulates A-Raf splicing in hepatocellular carcinoma by reducing production of a short dominant-negative A-Raf isoform and elevating full-length A-Raf transcript, thereby activating the Ras-MAPK-ERK pathway; hnRNP A2 knockdown inhibits ERK1/2 activation by EGF. siRNA knockdown of hnRNP A2; RT-PCR for A-Raf isoforms; ERK1/2 activation by EGF; overexpression studies; tumor formation assay RNA Medium 24572810
2015 ARAF mutations N217I and G322S in intrahepatic cholangiocarcinoma lead to activation of the MAPK pathway; N217I shows oncogenic potential in vitro (transformation assay). RNA/exome sequencing; MAPK pathway activation assay; in vitro transformation assay Nature communications Medium 25608663
2016 Subcellular localization of A-Raf determines its ability to inhibit MST2-mediated apoptosis: in proliferating and tumor cells, A-Raf localizes to mitochondria where it sequesters MST2; during epithelial differentiation, A-Raf re-localizes to the plasma membrane (regulated by KSR2 downregulation), releasing MST2 and rendering cells susceptible to apoptosis. Confocal immunofluorescence; subcellular fractionation; siRNA knockdown of KSR2; MCF7 differentiation model; apoptosis assays Cell death and differentiation Medium 26891695
2017 Alternative polyadenylation (APA) of Araf mRNA switches Araf protein isoforms during microglia activation, impacting production of downstream inflammatory cytokines. cTag-PAPERCLIP APA profiling; cell-type-specific protein isoform analysis; cytokine production assays in microglia Neuron Medium 28910620
2018 Hypusinated EIF5A1 regulates ARAF translation; ARAF mediates EIF5A1-dependent regulation of trophoblast migration and invasion via the integrin/ERK signaling pathway; ARAF levels are positively correlated with EIF5A1 in recurrent miscarriage villous tissues. EIF5A1 overexpression/knockdown; ARAF Western blot; trophoblast migration/invasion assays; villous explant culture; EIF5A1K50A hypusination mutant Cell death & disease Low 30206208
2019 The recurrent gain-of-function ARAF mutation S214P (loss of a conserved phosphorylation site) elevates ERK1/2 activity, enhances lymphangiogenic capacity, and causes disassembly of actin skeleton and VE-cadherin junctions in transduced cells; these effects are rescued by MEK inhibitor trametinib; an analogous lymphatic phenotype recreated in zebrafish is also rescued by MEK inhibition. Lentiviral transduction of ARAF-S214P; ERK1/2 phosphorylation assay; lymphangiogenesis assay; actin/VE-cadherin immunofluorescence; zebrafish morpholino model; trametinib rescue Nature medicine High 31263281
2020 ARAF-mediated resistance to the RAF dimer inhibitor belvarafenib requires both kinase activity and dimerization: ARAF kinase-domain mutations found in resistant melanoma cells and patient ctDNA form active dimers in the presence of belvarafenib; dimer-containing mutant ARAF is active under inhibitor conditions. Belvarafenib-resistant cell line generation; ctDNA sequencing from patients; ARAF mutant dimerization assays; kinase activity assays; panel of type II RAF inhibitors Nature High 33953400
2020 LXH254 (type II RAF inhibitor) preferentially inhibits BRAF and CRAF but largely spares ARAF; ARAF-mediated resistance to LXH254 requires both kinase function and dimerization; in RAS-mutant cells expressing only ARAF, LXH254 paradoxically activates MAPK signaling; loss of ARAF sensitizes RAS-mutant cells to LXH254. Biochemical kinase profiling; ARAF/BRAF/CRAF ablation (isogenic cell lines); kinase-impaired and dimer-deficient ARAF variants; paradoxical activation assays; xenograft models Clinical cancer research High 33355204
2022 ARAF activates RAS in a kinase-independent manner by binding to RAS and displacing the GTPase-activating protein NF1, thereby antagonizing NF1-mediated RAS-GTP hydrolysis, reducing ERK-dependent negative feedback on RAS, and increasing RAS-GTP levels; ARAF amplification in EGFR-mutant lung cancers confers resistance to EGFR inhibitors via this mechanism. Co-immunoprecipitation of ARAF-RAS-NF1 complexes; RAS-GTP pull-down; kinase-dead ARAF expression; ARAF overexpression/depletion; EGFR inhibitor resistance assays; combination with SHP2 inhibitor Molecular cell High 35613620
2022 CRAF:ARAF heterodimers are increased in KRAS-mutant cells (quantitative proteomics); depletion of both CRAF and ARAF rescues CRAF-loss phenotype; kinase-dead but not dimer-defective CRAF rescues growth, indicating that CRAF dimerization (including with ARAF) rather than kinase activity is required for maintaining appropriate MAPK signal intensity in KRAS-mutant tumors. Quantitative proteomics; siRNA depletion; rescue with kinase-dead and dimer-defective mutants; MAPK signaling assays; low-dose MEK/ERK inhibitor rescue Cell reports Medium 35139374
2022 ARAF suppresses ERBB3 expression in a kinase-independent manner by inhibiting ERBB3 promoter activity through the transcription factor KLF5; ARAF depletion increases ERBB3-AKT signaling and promotes metastasis; reconstitution of ARAF in ARAF-depleted cells reverses enhanced ERBB3-AKT signaling. ARAF siRNA depletion; ERBB3 promoter reporter assay; AKT signaling assays; KLF5 identification; rescue experiments Science advances Medium 35302851
2003 Trihydrophobin 1 (TH1) specifically binds to A-Raf kinase (but not B-Raf or C-Raf) both in vitro and in vivo; TH1 inhibits A-Raf kinase activity but does not affect B-Raf or C-Raf kinase activity; the A-Raf binding region of TH1 maps to amino acids 1–372; A-Raf kinase activity promotes binding to TH1. Co-immunoprecipitation; in vitro binding; Raf kinase assay; confocal colocalization; stable transfection with cell cycle analysis The Journal of biological chemistry Medium 14684750
2004 A-Raf associates with activated EGF receptor complexes and with PDGF receptor (PDGFR) complexes independently of prior PDGF treatment; expression of partially activated A-Raf mutant decreases tyrosine phosphorylation of PDGFR on Y857 and Y1021 (but not other docking sites) and alters PLCγ1 and PI3K activation. Co-immunoprecipitation; phospho-specific Western blotting of PDGFR tyrosines; A-Raf mutant overexpression; PLCγ1 and PI3K activity assays Cellular signalling Low 15763428

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2010 Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma. Nature 1433 20823850
2010 Acquired resistance to BRAF inhibitors mediated by a RAF kinase switch in melanoma can be overcome by cotargeting MEK and IGF-1R/PI3K. Cancer cell 1032 21156289
2015 Massive parallel sequencing uncovers actionable FGFR2-PPHLN1 fusion and ARAF mutations in intrahepatic cholangiocarcinoma. Nature communications 249 25608663
2009 A Raf-like MAPKKK gene DSM1 mediates drought resistance through reactive oxygen species scavenging in rice. Plant physiology 244 20007444
2003 Mutation analysis of the BRAF, ARAF and RAF-1 genes in human colorectal adenocarcinomas. Carcinogenesis 226 14688025
2011 A Raf-induced allosteric transition of KSR stimulates phosphorylation of MEK. Nature 211 21441910
2020 A RAF-SnRK2 kinase cascade mediates early osmotic stress signaling in higher plants. Nature communications 207 32001690
1995 Conditionally oncogenic forms of the A-Raf and B-Raf protein kinases display different biological and biochemical properties in NIH 3T3 cells. Molecular and cellular biology 198 7565795
2019 ARAF recurrent mutation causes central conducting lymphatic anomaly treatable with a MEK inhibitor. Nature medicine 162 31263281
1998 CNK, a RAF-binding multidomain protein required for RAS signaling. Cell 149 9814705
1987 The complete coding sequence of the human A-raf-1 oncogene and transforming activity of a human A-raf carrying retrovirus. Nucleic acids research 149 3029685
1995 Hypertrophic agonists stimulate the activities of the protein kinases c-Raf and A-Raf in cultured ventricular myocytes. The Journal of biological chemistry 143 7592840
1985 Rapid induction of hemopoietic neoplasms in newborn mice by a raf(mil)/myc recombinant murine retrovirus. Journal of virology 143 4009794
2014 Somatic activating ARAF mutations in Langerhans cell histiocytosis. Blood 140 24652991
2001 Nicotinic receptor-mediated activation by the tobacco-specific nitrosamine NNK of a Raf-1/MAP kinase pathway, resulting in phosphorylation of c-myc in human small cell lung carcinoma cells and pulmonary neuroendocrine cells. Journal of cancer research and clinical oncology 133 11768610
1996 Hepatitis B virus pX activates NF-kappa B-dependent transcription through a Raf-independent pathway. Journal of virology 129 8523586
1996 Post-natal lethality and neurological and gastrointestinal defects in mice with targeted disruption of the A-Raf protein kinase gene. Current biology : CB 125 8805280
2011 Increased leaf angle1, a Raf-like MAPKKK that interacts with a nuclear protein family, regulates mechanical tissue formation in the Lamina joint of rice. The Plant cell 115 22207574
1986 Characterization of murine A-raf, a new oncogene related to the v-raf oncogene. Molecular and cellular biology 114 3491291
1996 Selective activation of MEK1 but not MEK2 by A-Raf from epidermal growth factor-stimulated Hela cells. The Journal of biological chemistry 107 8621729
1988 In vitro-derived leukemic erythroid cell lines induced by a raf- and myc-containing retrovirus differentiate in response to erythropoietin. Proceedings of the National Academy of Sciences of the United States of America 105 2847163
2021 ARAF mutations confer resistance to the RAF inhibitor belvarafenib in melanoma. Nature 104 33953400
2014 Paradoxical activation of T cells via augmented ERK signaling mediated by a RAF inhibitor. Cancer immunology research 102 24416731
2004 A Raf/MEK/ERK signaling pathway is required for development of the sea urchin embryo micromere lineage through phosphorylation of the transcription factor Ets. Development (Cambridge, England) 100 14973284
1997 A raf-independent epidermal growth factor receptor autocrine loop is necessary for Ras transformation of rat intestinal epithelial cells. The Journal of biological chemistry 100 9228072
2014 Oncogenic and sorafenib-sensitive ARAF mutations in lung adenocarcinoma. The Journal of clinical investigation 99 24569458
2014 Splicing factor hnRNP A2 activates the Ras-MAPK-ERK pathway by controlling A-Raf splicing in hepatocellular carcinoma development. RNA (New York, N.Y.) 88 24572810
1997 A-Raf kinase is a new interacting partner of protein kinase CK2 beta subunit. FEBS letters 87 9042965
1985 Detection of a raf-related and two other transforming DNA sequences in human tumors maintained in nude mice. Proceedings of the National Academy of Sciences of the United States of America 79 2994056
2011 Synergistic action of a RAF inhibitor and a dual PI3K/mTOR inhibitor in thyroid cancer. Clinical cancer research : an official journal of the American Association for Cancer Research 78 21831957
1995 Insulin activates nuclear factor kappa B in mammalian cells through a Raf-1-mediated pathway. The Journal of biological chemistry 78 7592658
1997 The regulatory subunit of protein kinase CK2 is a specific A-Raf activator. FEBS letters 76 9042966
1997 Mutations of critical amino acids affect the biological and biochemical properties of oncogenic A-Raf and Raf-1. Oncogene 75 9285556
2020 LXH254, a Potent and Selective ARAF-Sparing Inhibitor of BRAF and CRAF for the Treatment of MAPK-Driven Tumors. Clinical cancer research : an official journal of the American Association for Cancer Research 74 33355204
2001 Crystal structures of YBHB and YBCL from Escherichia coli, two bacterial homologues to a Raf kinase inhibitor protein. Journal of molecular biology 70 11439028
1996 Activation of nuclear factor-kappaB via T cell receptor requires a Raf kinase and Ca2+ influx. Functional synergy between Raf and calcineurin. Journal of immunology (Baltimore, Md. : 1950) 70 8955173
2005 ZM336372, a Raf-1 activator, suppresses growth and neuroendocrine hormone levels in carcinoid tumor cells. Molecular cancer therapeutics 68 15956248
2016 Preclinical efficacy of a RAF inhibitor that evades paradoxical MAPK pathway activation in protein kinase BRAF-mutant lung cancer. Proceedings of the National Academy of Sciences of the United States of America 67 27834212
2010 Heterogeneous nuclear ribonucleoprotein H blocks MST2-mediated apoptosis in cancer cells by regulating A-Raf transcription. Cancer research 67 20145135
2020 Linc01232 promotes the metastasis of pancreatic cancer by suppressing the ubiquitin-mediated degradation of HNRNPA2B1 and activating the A-Raf-induced MAPK/ERK signaling pathway. Cancer letters 65 32814086
1999 A phosphatidylinositol 3-kinase-dependent pathway that differentially regulates c-Raf and A-Raf. The Journal of biological chemistry 65 10066754
2017 A Raf-like protein kinase BHP mediates blue light-dependent stomatal opening. Scientific reports 58 28358053
2006 Glycosylation of Pseudomonas aeruginosa strain Pa5196 type IV pilins with mycobacterium-like alpha-1,5-linked d-Araf oligosaccharides. Journal of bacteriology 58 17085575
2012 ARAF acts as a scaffold to stabilize BRAF:CRAF heterodimers. Oncogene 56 22926515
2004 Induction of apoptosis in human leukemia cells by the tyrosine kinase inhibitor adaphostin proceeds through a RAF-1/MEK/ERK- and AKT-dependent process. Oncogene 56 14647418
2000 Isoform-specific localization of A-RAF in mitochondria. Molecular and cellular biology 56 10848612
2017 cTag-PAPERCLIP Reveals Alternative Polyadenylation Promotes Cell-Type Specific Protein Diversity and Shifts Araf Isoforms with Microglia Activation. Neuron 55 28910620
2007 Bcl-2 protects endothelial cells against gamma-radiation via a Raf-MEK-ERK-survivin signaling pathway that is independent of cytochrome c release. Cancer research 55 17283155
2016 A Raf-like MAPKKK gene, GhRaf19, negatively regulates tolerance to drought and salt and positively regulates resistance to cold stress by modulating reactive oxygen species in cotton. Plant science : an international journal of experimental plant biology 52 27717463
2010 Blockade of the MEK/ERK pathway with a raf inhibitor prevents activation of pro-inflammatory mediators in cerebral arteries and reduction in cerebral blood flow after subarachnoid hemorrhage in a rat model. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 52 20424636
2011 c-Myc regulates RNA splicing of the A-Raf kinase and its activation of the ERK pathway. Cancer research 51 21512137
2002 ERK signalling and oncogene transformation are not impaired in cells lacking A-Raf. Oncogene 51 11821947
2020 KRASQ61H Preferentially Signals through MAPK in a RAF Dimer-Dependent Manner in Non-Small Cell Lung Cancer. Cancer research 50 32605999
2008 Modification of Pseudomonas aeruginosa Pa5196 type IV Pilins at multiple sites with D-Araf by a novel GT-C family Arabinosyltransferase, TfpW. Journal of bacteriology 47 18805982
2014 Dimerization of the kinase ARAF promotes MAPK pathway activation and cell migration. Science signaling 46 25097033
2016 PEBP1, a RAF kinase inhibitory protein, negatively regulates starvation-induced autophagy by direct interaction with LC3. Autophagy 45 27540684
2007 Regulation of pyruvate kinase type M2 by A-Raf: a possible glycolytic stop or go mechanism. Anticancer research 43 18225557
2003 A Raf-1 mutant that dissociates MEK/extracellular signal-regulated kinase activation from malignant transformation and differentiation but not proliferation. Molecular and cellular biology 43 12612072
2007 A-raf and B-raf are dispensable for normal endochondral bone development, and parathyroid hormone-related peptide suppresses extracellular signal-regulated kinase activation in hypertrophic chondrocytes. Molecular and cellular biology 41 17967876
1997 Selective inhibition of A-Raf and C-Raf mRNA expression by antisense oligodeoxynucleotides in rat vascular smooth muscle cells: role of A-Raf and C-Raf in serum-induced proliferation. Molecular pharmacology 41 9058592
2018 EIF5A1 promotes trophoblast migration and invasion via ARAF-mediated activation of the integrin/ERK signaling pathway. Cell death & disease 37 30206208
1995 The segment-specific pattern of A-raf expression in the mouse epididymis is regulated by testicular factors. Endocrinology 36 7750478
2022 CRAF dimerization with ARAF regulates KRAS-driven tumor growth. Cell reports 35 35139374
1992 Physical linkage of the A-raf-1, properdin, synapsin I, and TIMP genes on the human and mouse X chromosomes. Genomics 35 1572636
1986 Pks, a raf-related sequence in humans. Proceedings of the National Academy of Sciences of the United States of America 35 3529082
2005 Mutational analysis of the ARAF gene in human cancers. APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 34 15676015
2005 A-Raf and Raf-1 work together to influence transient ERK phosphorylation and Gl/S cell cycle progression. Oncogene 32 15856007
2008 Positive regulation of A-RAF by phosphorylation of isoform-specific hinge segment and identification of novel phosphorylation sites. The Journal of biological chemistry 31 18662992
2008 Sustained morphine treatment augments basal CGRP release from cultured primary sensory neurons in a Raf-1 dependent manner. European journal of pharmacology 30 18328477
2015 A-Raf: A new star of the family of raf kinases. Critical reviews in biochemistry and molecular biology 29 26508523
2013 Araf kinase antagonizes Nodal-Smad2 activity in mesendoderm development by directly phosphorylating the Smad2 linker region. Nature communications 29 23591895
2012 PKD controls mitotic Golgi complex fragmentation through a Raf-MEK1 pathway. Molecular biology of the cell 29 23242995
2007 Unique N-region determines low basal activity and limited inducibility of A-RAF kinase: the role of N-region in the evolutionary divergence of RAF kinase function in vertebrates. The Journal of biological chemistry 29 17613527
2020 Reversal of Cancer Multidrug Resistance (MDR) Mediated by ATP-Binding Cassette Transporter G2 (ABCG2) by AZ-628, a RAF Kinase Inhibitor. Frontiers in cell and developmental biology 28 33364237
2015 Activated k-ras, but not h-ras or N-ras, regulates brain neural stem cell proliferation in a raf/rb-dependent manner. Stem cells (Dayton, Ohio) 26 25788415
2000 Expression of the A-raf proto-oncogene in the normal adult and embryonic mouse. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research 26 10768864
2000 Using a phage display library to identify basic residues in A-Raf required to mediate binding to the Src homology 2 domains of the p85 subunit of phosphatidylinositol 3'-kinase. The Journal of biological chemistry 26 10967104
1991 A highly polymorphic dinucleotide repeat on the proximal short arm of the human X chromosome: linkage mapping of the synapsin I/A-raf-1 genes. American journal of human genetics 26 1905878
2022 A serine-rich effector from the stripe rust pathogen targets a Raf-like kinase to suppress host immunity. Plant physiology 25 35567492
1992 A Raf-1-related p110 polypeptide associates with the CD4-p56lck complex in T cells. Molecular and cellular biology 25 1406695
2022 TTYH3 Modulates Bladder Cancer Proliferation and Metastasis via FGFR1/H-Ras/A-Raf/MEK/ERK Pathway. International journal of molecular sciences 22 36142409
2004 Brain-derived neurotrophic factor-, epidermal growth factor-, or A-Raf-induced growth of HaCaT keratinocytes requires extracellular signal-regulated kinase. American journal of physiology. Cell physiology 21 15075211
2016 Differential localization of A-Raf regulates MST2-mediated apoptosis during epithelial differentiation. Cell death and differentiation 20 26891695
1993 Characterization of downstream elements in a Raf-1 pathway. Proceedings of the National Academy of Sciences of the United States of America 18 8430097
1987 A-raf oncogene localizes on mouse X chromosome to region some 10-17 centimorgans proximal to hypoxanthine phosphoribosyltransferase gene. Somatic cell and molecular genetics 18 3299748
1987 The complete primary structure of the rat A-raf cDNA coding region: conservation of the putative regulatory regions present in rat c-raf. Oncogene research 18 3449797
2022 ARAF protein kinase activates RAS by antagonizing its binding to RASGAP NF1. Molecular cell 17 35613620
2006 ZM336372, a Raf-1 activator, inhibits growth of pheochromocytoma cells. The Journal of surgical research 17 16603190
1996 Regulation of A-raf expression. Oncogene 17 8622887
1994 The complete sequence and promoter activity of the human A-raf-1 gene (ARAF1). Genomics 17 8020955
2019 Elevated Expression of A-Raf and FA2H in Hepatocellular Carcinoma is Associated with Lipid Metabolism Dysregulation and Cancer Progression. Anti-cancer agents in medicinal chemistry 16 30324893
2005 Identification of key residues in the A-Raf kinase important for phosphoinositide lipid binding specificity. Biochemistry 16 15736953
2003 Trihydrophobin 1 is a new negative regulator of A-Raf kinase. The Journal of biological chemistry 16 14684750
2009 A-RAF kinase functions in ARF6 regulated endocytic membrane traffic. PloS one 15 19247477
2019 α-Ketoglutaric Acid-Modified Carbonate Apatite Enhances Cellular Uptake and Cytotoxicity of a Raf- Kinase Inhibitor in Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways. Biomedicines 14 30609867
2006 Cortical migration defects in mice expressing A-RAF from the B-RAF locus. Molecular and cellular biology 14 16980614
2022 ARAF suppresses ERBB3 expression and metastasis in a subset of lung cancers. Science advances 13 35302851
2004 A-Raf associates with and regulates platelet-derived growth factor receptor signalling. Cellular signalling 13 15763428
1999 Identification, by cDNA microarray, of A-raf and proliferating cell nuclear antigen as genes induced in rat lung by exposure to diesel exhaust. Research communications in molecular pathology and pharmacology 13 10850371
2020 Silencing ARAF Suppresses the Malignant Phenotypes of Gallbladder Cancer Cells. BioMed research international 11 32923479

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