P11345 · RAF1_RAT
- ProteinRAF proto-oncogene serine/threonine-protein kinase
- GeneRaf1
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids648 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
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
Phosphorylates TNNT2/cardiac muscle troponin T
Catalytic activity
- ATP + L-seryl-[protein] = ADP + H+ + O-phospho-L-seryl-[protein]This reaction proceeds in the forward direction.
Cofactor
Note: Binds 2 Zn2+ ions per subunit.
Activity 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).
Features
Showing features for binding site, active site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 139 | Zn2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: H | ||||||
Binding site | 152 | Zn2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 155 | Zn2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 165 | Zn2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 168 | Zn2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 173 | Zn2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: H | ||||||
Binding site | 176 | Zn2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 184 | Zn2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: C | ||||||
Binding site | 355-363 | ATP (UniProtKB | ChEBI) | ||||
Sequence: IGSGSFGTV | ||||||
Binding site | 375 | ATP (UniProtKB | ChEBI) | ||||
Sequence: K | ||||||
Active site | 468 | Proton acceptor | ||||
Sequence: D |
GO annotations
Keywords
- Molecular function
- Ligand
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameRAF proto-oncogene serine/threonine-protein kinase
- EC number
- Alternative names
Gene names
Organism names
- Organism
- Taxonomic lineageEukaryota > Metazoa > Chordata > Craniata > Vertebrata > Euteleostomi > Mammalia > Eutheria > Euarchontoglires > Glires > Rodentia > Myomorpha > Muroidea > Muridae > Murinae > Rattus
Accessions
- Primary accessionP11345
Proteomes
Organism-specific databases
Subcellular Location
UniProt Annotation
GO Annotation
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).
Keywords
- Cellular component
Phenotypes & Variants
Keywords
- Disease
PTM/Processing
Features
Showing features for chain, modified residue.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Chain | PRO_0000086598 | 1-648 | RAF proto-oncogene serine/threonine-protein kinase | |||
Sequence: MEHIQGAWKTISNGFGLKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDSSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLQEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTASDSGVPAPPSFTMRRMRESVSRMPASSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPGSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQLQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSRLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF | ||||||
Modified residue | 29 | Phosphoserine; by MAPK1 | ||||
Sequence: S | ||||||
Modified residue | 43 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 233 | Phosphoserine; by PKA | ||||
Sequence: S | ||||||
Modified residue | 252 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 259 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 268 | Phosphothreonine; by autocatalysis | ||||
Sequence: T | ||||||
Modified residue | 269 | Phosphothreonine; by PKA | ||||
Sequence: T | ||||||
Modified residue | 289 | Phosphoserine; by MAPK1 | ||||
Sequence: S | ||||||
Modified residue | 296 | Phosphoserine; by MAPK1 | ||||
Sequence: S | ||||||
Modified residue | 301 | Phosphoserine; by MAPK1 | ||||
Sequence: S | ||||||
Modified residue | 338 | Phosphoserine; by PAK1, PAK2, PAK3 and PAK5 | ||||
Sequence: S | ||||||
Modified residue | 339 | Phosphoserine; by PAK1, PAK2 and PAK3 | ||||
Sequence: S | ||||||
Modified residue | 340 | Phosphotyrosine; by SRC | ||||
Sequence: Y | ||||||
Modified residue | 341 | Phosphotyrosine; by SRC | ||||
Sequence: Y | ||||||
Modified residue | 471 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 491 | Phosphothreonine | ||||
Sequence: T | ||||||
Modified residue | 494 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 497 | Phosphoserine; by PKC | ||||
Sequence: S | ||||||
Modified residue | 499 | Phosphoserine; by PKC | ||||
Sequence: S | ||||||
Modified residue | 563 | Symmetric dimethylarginine; by PRMT5 | ||||
Sequence: R | ||||||
Modified residue | 621 | Phosphoserine | ||||
Sequence: S | ||||||
Modified residue | 642 | Phosphoserine; by MAPK1 | ||||
Sequence: S |
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 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).
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).
Keywords
- PTM
Proteomic databases
PTM databases
Expression
Gene expression databases
Interaction
Subunit
Monomer (By similarity).
Homodimer (By similarity).
Heterodimerizes with BRAF and this heterodimer possesses a highly increased kinase activity compared to the respective homodimers or monomers (By similarity).
Heterodimerization is mitogen-regulated and enhanced by 14-3-3 proteins (By similarity).
MAPK1/ERK2 activation can induce a negative feedback that promotes the dissociation of the heterodimer (By similarity).
Forms a multiprotein complex with Ras (M-Ras/MRAS), SHOC2 and protein phosphatase 1 (PPP1CA, PPP1CB and PPP1CC) (By similarity).
Interacts with LZTR1 (By similarity).
Interacts with Ras proteins; the interaction is antagonized by RIN1 (By similarity).
Weakly interacts with RIT1 (By similarity).
Interacts with STK3/MST2; the interaction inhibits its pro-apoptotic activity (By similarity).
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 (By similarity).
Interacts with PAK1 (via kinase domain) (By similarity).
The phosphorylated form interacts with PIN1 (By similarity).
The Ser-338 and Ser-339 phosphorylated form (by PAK1) interacts with BCL2 (By similarity).
Interacts with PEBP1/RKIP and this interaction is enhanced if RAF1 is phosphorylated on residues Ser-338, Ser-339, Tyr-340 and Tyr-341 (By similarity).
Interacts with ADCY2, ADCY5, ADCY6, DGKH, RCAN1/DSCR1, PPP1R12A, PKB/AKT1, PPP2CA, PPP2R1B, SPRY2, SPRY4, CNKSR1/CNK1, KSR2 and PHB/prohibitin (By similarity).
Interacts with ROCK2 (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 (PubMed:19319189, PubMed:19878719).
Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones CDC37, PPP5C, TSC1 and client protein TSC2, CDK4, AKT, RAF1 and NR3C1; this complex does not contain co-chaperones STIP1/HOP and PTGES3/p23 (By similarity).
Interacts with MAP2K1/MEK1 and MAP2K2/MEK2 (PubMed:7565670).
In its active form, interacts with PRMT5 (By similarity).
Interacts with FAM83B; displaces 14-3-3 proteins from RAF1 and activates RAF1 (By similarity).
Interacts with PDE8A; the interaction promotes RAF1 activity (By similarity).
Interacts with MFHAS1 (By similarity).
Interacts with GLS (By similarity).
Interacts with YWHAZ. Interacts with NEK10 and MAP2K1; the interaction is direct with NEK10 and required for ERK1/2-signaling pathway activation in response to UV irradiation (By similarity).
Homodimer (By similarity).
Heterodimerizes with BRAF and this heterodimer possesses a highly increased kinase activity compared to the respective homodimers or monomers (By similarity).
Heterodimerization is mitogen-regulated and enhanced by 14-3-3 proteins (By similarity).
MAPK1/ERK2 activation can induce a negative feedback that promotes the dissociation of the heterodimer (By similarity).
Forms a multiprotein complex with Ras (M-Ras/MRAS), SHOC2 and protein phosphatase 1 (PPP1CA, PPP1CB and PPP1CC) (By similarity).
Interacts with LZTR1 (By similarity).
Interacts with Ras proteins; the interaction is antagonized by RIN1 (By similarity).
Weakly interacts with RIT1 (By similarity).
Interacts with STK3/MST2; the interaction inhibits its pro-apoptotic activity (By similarity).
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 (By similarity).
Interacts with PAK1 (via kinase domain) (By similarity).
The phosphorylated form interacts with PIN1 (By similarity).
The Ser-338 and Ser-339 phosphorylated form (by PAK1) interacts with BCL2 (By similarity).
Interacts with PEBP1/RKIP and this interaction is enhanced if RAF1 is phosphorylated on residues Ser-338, Ser-339, Tyr-340 and Tyr-341 (By similarity).
Interacts with ADCY2, ADCY5, ADCY6, DGKH, RCAN1/DSCR1, PPP1R12A, PKB/AKT1, PPP2CA, PPP2R1B, SPRY2, SPRY4, CNKSR1/CNK1, KSR2 and PHB/prohibitin (By similarity).
Interacts with ROCK2 (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 (PubMed:19319189, PubMed:19878719).
Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones CDC37, PPP5C, TSC1 and client protein TSC2, CDK4, AKT, RAF1 and NR3C1; this complex does not contain co-chaperones STIP1/HOP and PTGES3/p23 (By similarity).
Interacts with MAP2K1/MEK1 and MAP2K2/MEK2 (PubMed:7565670).
In its active form, interacts with PRMT5 (By similarity).
Interacts with FAM83B; displaces 14-3-3 proteins from RAF1 and activates RAF1 (By similarity).
Interacts with PDE8A; the interaction promotes RAF1 activity (By similarity).
Interacts with MFHAS1 (By similarity).
Interacts with GLS (By similarity).
Interacts with YWHAZ. Interacts with NEK10 and MAP2K1; the interaction is direct with NEK10 and required for ERK1/2-signaling pathway activation in response to UV irradiation (By similarity).
Binary interactions
Type | Entry 1 | Entry 2 | Number of experiments | Intact | |
---|---|---|---|---|---|
BINARY | P11345 | Arrb1 P29066 | 5 | EBI-931534, EBI-4303019 |
Protein-protein interaction databases
Structure
Family & Domains
Features
Showing features for domain, zinc finger, region, compositional bias.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Domain | 56-131 | RBD | ||||
Sequence: NTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLQEHKGKKARLDWNTDAASLIGEELQVDFL | ||||||
Zinc finger | 138-184 | Phorbol-ester/DAG-type | ||||
Sequence: THNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMC | ||||||
Region | 217-335 | Disordered | ||||
Sequence: MRESVSRMPASSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPGSGTQEKNKIRPRGQ | ||||||
Compositional bias | 222-269 | Polar residues | ||||
Sequence: SRMPASSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTT | ||||||
Compositional bias | 284-310 | Polar residues | ||||
Sequence: HSESASPSALSSSPNNLSPTGWSQPKT | ||||||
Region | 331-349 | Interaction with PEBP1/RKIP | ||||
Sequence: RPRGQRDSSYYWEIEASEV | ||||||
Domain | 349-609 | Protein kinase | ||||
Sequence: VMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQLQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSRLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELL |
Sequence similarities
Keywords
- Domain
Phylogenomic databases
Family and domain databases
Sequence
- Sequence statusComplete
- Length648
- Mass (Da)72,928
- Last updated1989-07-01 v1
- ChecksumE9AFB5975064193E
Computationally mapped potential isoform sequences
There are 3 potential isoforms mapped to this entry
Entry | Entry name | Gene name | Length | ||
---|---|---|---|---|---|
A0A8I6ALJ3 | A0A8I6ALJ3_RAT | Raf1 | 608 | ||
A0A8L2Q6F4 | A0A8L2Q6F4_RAT | Raf1 | 646 | ||
A0A8I5ZWZ7 | A0A8I5ZWZ7_RAT | Raf1 | 668 |
Features
Showing features for compositional bias.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Compositional bias | 222-269 | Polar residues | ||||
Sequence: SRMPASSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTT | ||||||
Compositional bias | 284-310 | Polar residues | ||||
Sequence: HSESASPSALSSSPNNLSPTGWSQPKT |
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
M15427 EMBL· GenBank· DDBJ | AAA42001.1 EMBL· GenBank· DDBJ | mRNA | ||
BC062071 EMBL· GenBank· DDBJ | AAH62071.1 EMBL· GenBank· DDBJ | mRNA |