Ras (Ha-Ras, Ki-Ras, N-Ras) is implicated in the regulation of various
cell functions such as gene expression and cell proliferation downstr
eam from specific extracellular signals. Here, we partially purified a
Ras-inter acting protein with molecular mass of about 180 kDa (p180)
from bovine brain membrane extract by glutathione S-transferase (GST)
Ha-Ras affinity column chromatography. This protein bound to the GTP g
amma S (guanosine 5'-(3-O-thio)triphosphate, a nonhydrolyzable GTP ana
log). GST-Ha-Ras affinity column but not to those containing GDP . GST
-Ha-Ras or GTP gamma S . GST-Ha-Ras with a mutation in the effector do
main (Ha-Ras(A38)). The amino acid sequences of the peptides derived f
rom p180 were almost identical to those of human AF-6 that is identifi
ed as the fusion partner of the ALL-1 protein. The ALL-1/AF-6 chimeric
protein is the critical product of the t (6:11) abnormality associate
d with some human leukemia. AF-6 has a GLGF/Dlg homology repeat (DHR)
motif and shows a high degree of sequence similarity with Drosophila C
anoe, which is assumed to function downstream from Notch in a common d
evelopmental pathway. The recombinant N-terminal domain of AF-6 and Ca
noe specifically interacted with GTP gamma S . GST-Ha-Ras. The known R
as target c-Raf-1 inhibited the interaction of AF 6 with GTP gamma S .
GST-Ha-Ras. These results indicate that AF-6 and Canoe are putative t
argets for Ras.