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TitleStructural insights into the BRAF monomer-to-dimer transition mediated by RAS binding.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 486, Year 2022
Publish dateJan 25, 2022
AuthorsJuliana A Martinez Fiesco / David E Durrant / Deborah K Morrison / Ping Zhang /
PubMed AbstractRAF kinases are essential effectors of RAS, but how RAS binding initiates the conformational changes needed for autoinhibited RAF monomers to form active dimers has remained unclear. Here, we present ...RAF kinases are essential effectors of RAS, but how RAS binding initiates the conformational changes needed for autoinhibited RAF monomers to form active dimers has remained unclear. Here, we present cryo-electron microscopy structures of full-length BRAF complexes derived from mammalian cells: autoinhibited, monomeric BRAF:14-3-3:MEK and BRAF:14-3-3 complexes, and an inhibitor-bound, dimeric BRAF:14-3-3 complex, at 3.7, 4.1, and 3.9 Å resolution, respectively. In both autoinhibited, monomeric structures, the RAS binding domain (RBD) of BRAF is resolved, revealing that the RBD forms an extensive contact interface with the 14-3-3 protomer bound to the BRAF C-terminal site and that key basic residues required for RBD-RAS binding are exposed. Moreover, through structure-guided mutational studies, our findings indicate that RAS-RAF binding is a dynamic process and that RBD residues at the center of the RBD:14-3-3 interface have a dual function, first contributing to RAF autoinhibition and then to the full spectrum of RAS-RBD interactions.
External linksNat Commun / PubMed:35078985 / PubMed Central
MethodsEM (single particle)
Resolution3.66 - 4.07 Å
Structure data

23813

7mfd

EMDB-23813, PDB-7mfd:
Autoinhibited BRAF:(14-3-3)2:MEK complex with the BRAF RBD resolved
Method: EM (single particle) / Resolution: 3.66 Å

23814

7mfe

EMDB-23814, PDB-7mfe:
Autoinhibited BRAF:(14-3-3)2 complex with the BRAF RBD resolved
Method: EM (single particle) / Resolution: 4.07 Å

23815

7mff

EMDB-23815, PDB-7mff:
Dimeric (BRAF)2:(14-3-3)2 complex bound to SB590885 Inhibitor
Method: EM (single particle) / Resolution: 3.89 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-CHU:
N-(3-fluoro-4-{[4-methyl-2-oxo-7-(pyrimidin-2-yloxy)-2H-chromen-3-yl]methyl}pyridin-2-yl)-N'-methylsulfuric diamide

ChemComp-215:
(1Z)-5-(2-{4-[2-(DIMETHYLAMINO)ETHOXY]PHENYL}-5-PYRIDIN-4-YL-1H-IMIDAZOL-4-YL)INDAN-1-ONE OXIME

Source
  • homo sapiens (human)
  • Human (human)
KeywordsSIGNALING PROTEIN/TRANSFERASE / B-Raf / MEK / 14-3-3 / B-Raf complex / B-Raf monomer / Inactive B-Raf / Serine/threonine-protein kinase B-raf / RBD / signaling protein / SIGNALING PROTEIN-TRANSFERASE complex / B-Raf dimer / Active B-Raf / SB590885

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