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P11345 (RAF1_RAT) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 150. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
RAF proto-oncogene serine/threonine-protein kinase

EC=2.7.11.1
Alternative name(s):
Proto-oncogene c-RAF
Short name=cRaf
Raf-1
Gene names
Name:Raf1
Synonyms:Raf
OrganismRattus norvegicus (Rat) [Reference proteome]
Taxonomic identifier10116 [NCBI]
Taxonomic lineageEukaryotaMetazoaChordataCraniataVertebrataEuteleostomiMammaliaEutheriaEuarchontogliresGliresRodentiaSciurognathiMuroideaMuridaeMurinaeRattus

Protein attributes

Sequence length648 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation By similarity. Phosphorylates TNNT2/cardiac muscle troponin T. Ref.3 Ref.6

Catalytic activity

ATP + a protein = ADP + a phosphoprotein.

Cofactor

Binds 2 zinc ions per subunit By similarity.

Enzyme regulation

Regulation is a highly complex process involving membrane recruitment, protein-protein interactions, dimerization, and phosphorylation/dephosphorylation events. Ras-GTP recruits RAF1 to the membrane, thereby promoting its activation. The inactive conformation of RAF1 is maintained by autoinhibitory interactions occurring between the N-terminal regulatory and the C-terminal catalytic domains and by the binding of a 14-3-3 protein that contacts two phosphorylation sites, Ser-259 and Ser-621. Upon mitogenic stimulation, Ras and PPP2R1A cooperate to release autoinhibition and the subsequent phosphorylation of activating sites: Ser-338, Tyr-341, Thr-491, and Ser-494, yields a fully active kinase. Through a negative feedback mechanism involving MAPK1/ERK2, RAF1 is phosphorylated on Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2, which yields an inactive, desensitized kinase. The signaling-competent conformation of RAF1 is finally re-established by the coordinated action of PIN1, a prolyl isomerase that converts pSer and pThr residues from the cis to the trans conformation, which is preferentially recognized and dephosphorylated by PPP2R1A. Activated by homodimerization and heterodimerization (with BRAF). Also regulated through association with other proteins such as KSR2, CNKSR1/CNK1, PEBP1/RKIP, PHB/prohibitin and SPRY4. PEBP1/RKIP acts by dissociating RAF1 from its substrates MAP2K1/MEK1 and MAP2K2/MEK2. PHB/prohibitin facilitates the displacement of 14-3-3 from RAF1 by activated Ras, thereby promoting cell membrane localization and phosphorylation of RAF1 at the activating Ser-338. SPRY4 inhibits Ras-independent, but not Ras-dependent, activation of RAF1. CNKSR1/CNK1 regulates Src-mediated RAF1 activation By similarity.

Subunit structure

Monomer. Homodimer. Heterodimerizes with BRAF and this heterodimer possesses a highly increased kinase activity compared to the respective homodimers or monomers. Heterodimerization is mitogen-regulated and enhanced by 14-3-3 proteins. MAPK1/ERK2 activation can induce a negative feedback that promotes the dissociation of the heterodimer. Forms a multiprotein complex with Ras (M-Ras/MRAS), SHOC2 and protein phosphatase 1 (PPP1CA, PPP1CB and PPP1CC). Interacts with Ras proteins; the interaction is antagonized by RIN1. Weakly interacts with RIT1 By similarity. Interacts with STK3/MST2; the interaction inhibits its pro-apoptotic activity. Interacts (when phosphorylated at Ser-259) with YWHAZ (unphosphorylated at 'Thr-232') By similarity. Interacts with MAP3K5/ASF1 (via N-terminus) and this interaction inhibits the proapoptotic function of MAP3K5/ASK1. Interacts with PAK1 (via kinase domain). The phosphorylated form interacts with PIN1. The Ser-338 and Ser-339 phosphorylated form (by PAK1) interacts with BCL2. Interacts with PEBP1/RKIP and this interaction is enhanced if RAF1 is phosphorylated on residues Ser-338, Ser-339, Tyr-340 and Tyr-341. Interacts with ADCY2, ADCY5, ADCY6, DGKH, RCAN1/DSCR1, ROCK2, PPP1R12A, PKB/AKT1, PPP2CA, PPP2R1B, SPRY2, SPRY4, CNKSR1/CNK1, KSR2 and PHB/prohibitin By similarity. Interacts (via N-terminus) with RGS14 (via RBD domains); the interaction mediates the formation of a ternary complex with BRAF, a ternary complex inhibited by GNAI1. Interacts with MAP2K1/MEK1 and MAP2K2/MEK2. In its active form, interacts with PRMT5 By similarity. Ref.4 Ref.5 Ref.6

Subcellular location

Cytoplasm By similarity. Cell membrane By similarity. Mitochondrion By similarity. Nucleus By similarity. Note: Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. Phosphorylation at Ser-259 impairs its membrane accumulation. Recruited to the cell membrane by the active Ras protein. Phosphorylation at Ser-338 and Ser-339 by PAK1 is required for its mitochondrial localization. Retinoic acid-induced Ser-621 phosphorylated form of RAF1 is predominantly localized at the nucleus By similarity. Ref.5

Post-translational modification

Phosphorylation at Thr-269, Ser-338, Tyr-341, Thr-491 and Ser-494 results in its activation. Phosphorylation at Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2 results in its inactivation. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity. Phosphorylation at Ser-338 by PAK1 and PAK7/PAK5 and Ser-339 by PAK1 is required for its mitochondrial localization By similarity. Phosphorylation at Ser-621 in response to growth factor treatment stabilizes the protein, possibly by preventing proteasomal degradation. Phosphorylation at Ser-289, Ser-296, Ser-301, Ser-338 and Ser-621 are somehow linked to the methylation potential of cells. Treatment of cells with HGF in the presence of the methylation inhibitor 5'-methylthioadenosine (MTA) results in increased phosphorylation at Ser-338 and Ser-621 and decreased phosphorylation at Ser-296, Ser-301 and Ser-338. Dephosphorylation at SER-338 by PPP5C results in a decreased of activity By similarity.

Methylated in response to EGF treatment. This modification leads to destabilization of the protein, possibly through proteasomal degradation By similarity. Ref.7

Sequence similarities

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.

Contains 1 phorbol-ester/DAG-type zinc finger.

Contains 1 protein kinase domain.

Contains 1 RBD (Ras-binding) domain.

Ontologies

Keywords
   Cellular componentCell membrane
Cytoplasm
Membrane
Mitochondrion
Nucleus
   DiseaseProto-oncogene
   DomainZinc-finger
   LigandATP-binding
Metal-binding
Nucleotide-binding
Zinc
   Molecular functionKinase
Serine/threonine-protein kinase
Transferase
   PTMMethylation
Phosphoprotein
   Technical term3D-structure
Complete proteome
Reference proteome
Gene Ontology (GO)
   Biological_processMAPK cascade

Inferred from direct assay PubMed 15886202. Source: RGD

activation of MAPKK activity

Inferred from direct assay PubMed 10648842PubMed 16272159PubMed 8603510. Source: GOC

death-inducing signaling complex assembly

Inferred from electronic annotation. Source: Ensembl

heart development

Inferred from expression pattern PubMed 9779826. Source: RGD

intermediate filament cytoskeleton organization

Inferred from electronic annotation. Source: Ensembl

intracellular signal transduction

Traceable author statement PubMed 9779826. Source: RGD

negative regulation of apoptotic process

Inferred from electronic annotation. Source: Ensembl

negative regulation of cell proliferation

Inferred from electronic annotation. Source: Ensembl

negative regulation of extrinsic apoptotic signaling pathway via death domain receptors

Inferred from electronic annotation. Source: Ensembl

negative regulation of protein complex assembly

Inferred from electronic annotation. Source: Ensembl

neurotrophin TRK receptor signaling pathway

Inferred from electronic annotation. Source: Ensembl

positive regulation of peptidyl-serine phosphorylation

Inferred from electronic annotation. Source: Ensembl

protein phosphorylation

Inferred from direct assay PubMed 10648842PubMed 16272159. Source: RGD

response to hypoxia

Inferred from direct assay PubMed 8603510. Source: RGD

   Cellular_componentcytoplasm

Inferred from direct assay Ref.5. Source: UniProtKB

cytosol

Traceable author statement PubMed 9779826. Source: RGD

mitochondrion

Inferred from electronic annotation. Source: UniProtKB-SubCell

nucleus

Inferred from electronic annotation. Source: UniProtKB-SubCell

plasma membrane

Inferred from direct assay Ref.5. Source: UniProtKB

pseudopodium

Inferred from electronic annotation. Source: Ensembl

   Molecular_functionATP binding

Inferred from direct assay PubMed 16272159. Source: RGD

MAP kinase kinase kinase activity

Inferred from direct assay PubMed 10648842PubMed 16272159PubMed 8603510. Source: RGD

metal ion binding

Inferred from electronic annotation. Source: UniProtKB-KW

mitogen-activated protein kinase kinase binding

Inferred from mutant phenotype PubMed 16508002. Source: RGD

protein heterodimerization activity

Inferred from physical interaction PubMed 16508002. Source: RGD

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 648648RAF proto-oncogene serine/threonine-protein kinase
PRO_0000086598

Regions

Domain56 – 13176RBD
Domain349 – 609261Protein kinase
Zinc finger138 – 18447Phorbol-ester/DAG-type
Nucleotide binding355 – 3639ATP By similarity
Region331 – 34919Interaction with PEBP1/RKIP By similarity

Sites

Active site4681Proton acceptor By similarity
Metal binding1391Zinc 1 By similarity
Metal binding1521Zinc 2 By similarity
Metal binding1551Zinc 2 By similarity
Metal binding1651Zinc 1 By similarity
Metal binding1681Zinc 1 By similarity
Metal binding1731Zinc 2 By similarity
Metal binding1761Zinc 2 By similarity
Metal binding1841Zinc 1 By similarity
Binding site3751ATP By similarity

Amino acid modifications

Modified residue291Phosphoserine; by MAPK1 By similarity
Modified residue431Phosphoserine; by PKA and MAPK1 By similarity
Modified residue2331Phosphoserine; by PKA By similarity
Modified residue2521Phosphoserine By similarity
Modified residue2591Phosphoserine; by PKA, PKC and PKB/AKT1 By similarity
Modified residue2681Phosphothreonine; by autocatalysis By similarity
Modified residue2691Phosphothreonine; by PKA By similarity
Modified residue2891Phosphoserine; by MAPK1 By similarity
Modified residue2961Phosphoserine; by MAPK1 By similarity
Modified residue3011Phosphoserine; by MAPK1 By similarity
Modified residue3381Phosphoserine; by PAK1, PAK2, PAK3 and PAK7/PAK5 By similarity
Modified residue3391Phosphoserine; by PAK1, PAK2 and PAK3 By similarity
Modified residue3401Phosphotyrosine; by SRC By similarity
Modified residue3411Phosphotyrosine; by SRC By similarity
Modified residue4711Phosphoserine By similarity
Modified residue4911Phosphothreonine By similarity
Modified residue4941Phosphoserine By similarity
Modified residue4971Phosphoserine; by PKC By similarity
Modified residue4991Phosphoserine; by PKC By similarity
Modified residue5631Symmetric dimethylarginine; by PRMT5 By similarity
Modified residue6211Phosphoserine By similarity
Modified residue6421Phosphoserine; by MAPK1 By similarity

Secondary structure

.................. 648
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P11345 [UniParc].

Last modified July 1, 1989. Version 1.
Checksum: E9AFB5975064193E

FASTA64872,928
        10         20         30         40         50         60 
MEHIQGAWKT ISNGFGLKDA VFDGSSCISP TIVQQFGYQR RASDDGKLTD SSKTSNTIRV 

        70         80         90        100        110        120 
FLPNKQRTVV NVRNGMSLHD CLMKALKVRG LQPECCAVFR LLQEHKGKKA RLDWNTDAAS 

       130        140        150        160        170        180 
LIGEELQVDF LDHVPLTTHN FARKTFLKLA FCDICQKFLL NGFRCQTCGY KFHEHCSTKV 

       190        200        210        220        230        240 
PTMCVDWSNI RQLLLFPNST ASDSGVPAPP SFTMRRMRES VSRMPASSQH RYSTPHAFTF 

       250        260        270        280        290        300 
NTSSPSSEGS LSQRQRSTST PNVHMVSTTL PVDSRMIEDA IRSHSESASP SALSSSPNNL 

       310        320        330        340        350        360 
SPTGWSQPKT PVPAQRERAP GSGTQEKNKI RPRGQRDSSY YWEIEASEVM LSTRIGSGSF 

       370        380        390        400        410        420 
GTVYKGKWHG DVAVKILKVV DPTPEQLQAF RNEVAVLRKT RHVNILLFMG YMTKDNLAIV 

       430        440        450        460        470        480 
TQWCEGSSLY KHLHVQETKF QMFQLIDIAR QTAQGMDYLH AKNIIHRDMK SNNIFLHEGL 

       490        500        510        520        530        540 
TVKIGDFGLA TVKSRWSGSQ QVEQPTGSVL WMAPEVIRMQ DNNPFSFQSD VYSYGIVLYE 

       550        560        570        580        590        600 
LMTGELPYSH INNRDQIIFM VGRGYASPDL SRLYKNCPKA MKRLVADCVK KVKEERPLFP 

       610        620        630        640 
QILSSIELLQ HSLPKINRSA SEPSLHRAAH TEDINACTLT TSPRLPVF 

« Hide

References

« Hide 'large scale' references
[1]"Rat c-raf oncogene activation by a rearrangement that produces a fused protein."
Ishikawa F., Takaku F., Nagao M., Sugimura T.
Mol. Cell. Biol. 7:1226-1232(1987) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[2]"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC)."
The MGC Project Team
Genome Res. 14:2121-2127(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
Tissue: Prostate.
[3]"Raf-1: a novel cardiac troponin T kinase."
Pfleiderer P., Sumandea M.P., Rybin V.O., Wang C., Steinberg S.F.
J. Muscle Res. Cell Motil. 30:67-72(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN PHOSPHORYLATION OF TNNT2.
[4]"Regulator of G-protein signaling 14 (RGS14) is a selective H-Ras effector."
Willard F.S., Willard M.D., Kimple A.J., Soundararajan M., Oestreich E.A., Li X., Sowa N.A., Kimple R.J., Doyle D.A., Der C.J., Zylka M.J., Snider W.D., Siderovski D.P.
PLoS ONE 4:E4884-E4884(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH RGS14.
[5]"RGS14 is a multifunctional scaffold that integrates G protein and Ras/Raf MAPkinase signalling pathways."
Shu F.J., Ramineni S., Hepler J.R.
Cell. Signal. 22:366-376(2010) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH RGS14, SUBCELLULAR LOCATION.
[6]"A proline-rich sequence unique to MEK1 and MEK2 is required for raf binding and regulates MEK function."
Catling A.D., Schaeffer H.J., Reuter C.W., Reddy G.R., Weber M.J.
Mol. Cell. Biol. 15:5214-5225(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH MAP2K1/MEK1 AND MAP2K2/MEK2, FUNCTION.
[7]"Protein arginine methyltransferase 5 regulates ERK1/2 signal transduction amplitude and cell fate through CRAF."
Andreu-Perez P., Esteve-Puig R., de Torre-Minguela C., Lopez-Fauqued M., Bech-Serra J.J., Tenbaum S., Garcia-Trevijano E.R., Canals F., Merlino G., Avila M.A., Recio J.A.
Sci. Signal. 4:RA58-RA58(2011) [PubMed] [Europe PMC] [Abstract]
Cited for: METHYLATION.
[8]"Nuclear magnetic resonance and molecular dynamics studies on the interactions of the Ras-binding domain of Raf-1 with wild-type and mutant Ras proteins."
Terada T., Ito Y., Shirouzu M., Tateno M., Hashimoto K., Kigawa T., Ebisuzaki T., Takio K., Shibata T., Yokoyama S., Smith B.O., Laue E.D., Cooper J.A.
J. Mol. Biol. 286:219-232(1999) [PubMed] [Europe PMC] [Abstract]
Cited for: STRUCTURE BY NMR OF 51-131.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
M15427 mRNA. Translation: AAA42001.1.
BC062071 mRNA. Translation: AAH62071.1.
PIRTVRTRF. A26126.
RefSeqNP_036771.1. NM_012639.2.
UniGeneRn.33262.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1RRBNMR-A51-131[»]
ProteinModelPortalP11345.
SMRP11345. Positions 56-131, 136-187, 342-615.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

BioGrid246833. 7 interactions.
DIPDIP-1073N.
IntActP11345. 1 interaction.
STRING10116.ENSRNOP00000061173.

PTM databases

PhosphoSiteP11345.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

GeneID24703.
KEGGrno:24703.
UCSCRGD:3531. rat.

Organism-specific databases

CTD5894.
RGD3531. Raf1.

Phylogenomic databases

eggNOGCOG0515.
HOGENOMHOG000252972.
HOVERGENHBG001886.
InParanoidP11345.
KOK04366.

Enzyme and pathway databases

BRENDA2.7.10.2. 5301.

Gene expression databases

ArrayExpressP11345.
GenevestigatorP11345.

Family and domain databases

InterProIPR020454. DAG/PE-bd.
IPR011009. Kinase-like_dom.
IPR002219. Prot_Kinase_C-like_PE/DAG-bd.
IPR000719. Prot_kinase_dom.
IPR017441. Protein_kinase_ATP_BS.
IPR003116. Raf-like_ras-bd.
IPR001245. Ser-Thr/Tyr_kinase_cat_dom.
IPR008271. Ser/Thr_kinase_AS.
[Graphical view]
PfamPF00130. C1_1. 1 hit.
PF07714. Pkinase_Tyr. 1 hit.
PF02196. RBD. 1 hit.
[Graphical view]
PRINTSPR00008. DAGPEDOMAIN.
SMARTSM00109. C1. 1 hit.
SM00455. RBD. 1 hit.
[Graphical view]
SUPFAMSSF56112. SSF56112. 1 hit.
PROSITEPS00107. PROTEIN_KINASE_ATP. 1 hit.
PS50011. PROTEIN_KINASE_DOM. 1 hit.
PS00108. PROTEIN_KINASE_ST. 1 hit.
PS50898. RBD. 1 hit.
PS00479. ZF_DAG_PE_1. 1 hit.
PS50081. ZF_DAG_PE_2. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP11345.
NextBio604169.
PROP11345.

Entry information

Entry nameRAF1_RAT
AccessionPrimary (citable) accession number: P11345
Entry history
Integrated into UniProtKB/Swiss-Prot: July 1, 1989
Last sequence update: July 1, 1989
Last modified: April 16, 2014
This is version 150 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programChordata Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references