Identification of AF-6 and canoe as putative targets for Ras.
Ras (Ha-Ras, Ki-Ras, N-Ras) is implicated in the regulation of various cell functions such as gene expression and cell proliferation downstream from specific extracellular signals. Here, we partially purified a Ras-interacting 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 gamma S (guanosine 5'-(3-O-thio)triphosphate, a nonhydrolyzable GTP analog).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 domain (Ha-RasA38). The amino acid sequences of the peptides derived from p180 were almost identical to those of human AF-6 that is identified 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 associated with some human leukemia. AF-6 has a GLGF/Dlg homology repeat (DHR) motif and shows a high degree of sequence similarity with Drosophila Canoe, which is assumed to function downstream from Notch in a common developmental pathway. The recombinant N-terminal domain of AF-6 and Canoe specifically interacted with GTP gamma S.GST-Ha-Ras. The known Ras 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 targets for Ras.
Kuriyama
M
M
Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma, Japan.
Harada
N
N
Kuroda
S
S
Yamamoto
T
T
Nakafuku
M
M
Iwamatsu
A
A
Yamamoto
D
D
Prasad
R
R
Croce
C
C
Canaani
E
E
Kaibuchi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D029721:Drosophila Proteins; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D008969:Molecular Sequence Data; D009218:Myosins / Q000378:metabolism; D015689:Oncogene Protein p21(ras) / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017421:Sequence Analysis
C095516:AFDN protein, human; D029721:Drosophila Proteins; D011506:Proteins; C092747:cno protein, Drosophila; D009218:Myosins; D016547:Kinesins; D015689:Oncogene Protein p21(ras)