Identification of AF-6 and Canoe as Putative Targets for Ras (*)
- Masamitsu Kuriyama(1),
- Naozumi Harada(1),
- Shinya Kuroda(1),
- Takaharu Yamamoto(1),
- Masato Nakafuku(1),
- Akihiro Iwamatsu(2),
- Daisuke Yamamoto(3),
- Raj Prasad(4),
- Carlo Croce(4),
- Eli Canaani(5) and
- Kozo Kaibuchi(1)(§)
- From the (1)Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-01, Japan
- (2)Central Laboratories for Key Technology, Kirin Brewery Co. Ltd., Yokohama 236, Japan
- (3)Mitsubishi Kasei Institute of Life Sciences and ERATO Yamamoto Behavior Genes Project, Machida 194, Japan
- (4)Thomas Jefferson University, Philadelphia, Pennsylvania 19107, and
- (5)Weizmann Institute of Science, Rehovot 76100, Israel
- § To whom correspondence should be addressed. Tel.: 81-7437-2-5440; Fax: 81-7437-2-5449; kaibuchi{at}bs.aist-nara.ac.jp.
Abstract
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
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
S•GST-Ha-Ras with a mutation in the effector domain (Ha-Ras
). 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
S•GST-Ha-Ras. The known Ras target c-Raf-1 inhibited the interaction of AF-6 with GTP
S•GST-Ha-Ras. These results indicate that AF-6 and Canoe are putative targets for Ras.
Footnotes
-
↵* This study was supported by grants-in-aid for Scientific Research and for Cancer Research from the Ministry of Education, Science, and Culture, Japan(1995) and by a grant from the Yamanouchi Foundation for Research on Metabolic Disease(1995). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
-
↵1 The abbreviations used are:
- MAP
-
mitogen-activated protein
- Ral GDS
-
Ral guanine nucleotide dissociation stimulator
- GST
-
glutathione S-transferase
- AF-6
-
ALL-1 fusion partner from chromosome 6
- ALL
-
acute lymphoblastic leukemia
- GTP
S -
guanosine 5′-(3-O-thio)triphosphate
- aa
-
amino acids
- MBP
-
maltose-binding protein
- G proteins
-
GTP-binding proteins
- PAGE
-
polyacrylamide gel electrophoresis.
-
- Received October 10, 1995.
- Revision received November 7, 1995.
- © 1996 by The American Society for Biochemistry and Molecular Biology, Inc.
