Mutations of the BRAF gene in human cancer

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Davies, H. ; Bignell, G.R. ; Cox, C. ; Stephens, P. ; Edkins, S. ; Clegg, S. ; Teague, J. ; Woffendin, H. ; Garnett, M.J. ; Bottomley, W. ; Davis, N. ; Dicks, E. ; Ewing, R. ; Floyd, Y. ; Gray, K. ; Hall, S. ; Hawes, R. ; Hughes, J. ; Kosmidou, V. ; Menzies, A. ; Mould, C. ; Parker, A. ; Stevens, C. ; Watt, S. ; Hooper, S. ; Wilson, R. ; Jayatilake, H. ; Gusterson, B.A. ; Cooper, C. ; Shipley, J. ... view all 52 authors (2002)

Cancers arise owing to the accumulation of mutations in critical genes that alter normal programmes of cell proliferation, differentiation and death. As the first stage of a systematic genome-wide screen for these genes, we have prioritized for analysis signalling pathways in which at least one gene is mutated in human cancer. The RAS RAF MEK ERK MAP kinase pathway mediates cellular responses to growth signals. RAS is mutated to an oncogenic form in about 15% of human cancer. The three RAF genes code for cytoplasmic serine/threonine kinases that are regulated by binding RAS. Here we report BRAF somatic missense mutations in 66% of malignant melanomas and at lower frequency in a wide range of human cancers. All mutations are within the kinase domain, with a single substitution (V599E) accounting for 80%. Mutated BRAF proteins have elevated kinase activity and are transforming in NIH3T3 cells. Furthermore, RAS function is not required for the growth of cancer cell lines with the V599E mutation. As BRAF is a serine/threonine kinase that is commonly activated by somatic point mutation in human cancer, it may provide new therapeutic opportunities in malignant melanoma.
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    Allele Transformed foci per mg DNA Fold increase over wild-type BRAF ............................................................................................................................................................................. WTBRAF 1.3 - V599E 180 138 £ DAVE 0 - L596V 90 70 £ DALV 0 - G463V 130 100 £ GG1426VH8ARAS 129,0000 9,62900££ ............................................................................................................................................................................. NIH3T3 cells were transfected as described in Methods. Transformed foci contained cells like Rasor Raf1-transformed cells-which are refractile and frequently bipolar-and often contained the giant cells typical of RAS or RAF1 transformation. DAVE and DALV are kinase-inactive versions of V599E and L596V, respectively, in which D593 of the conserved DFG motif is replaced by alanine to generate a kinase-dead variant.

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