Characterization of full-length neurofibromin: tubulin inhibits Ras GAP activity.
Full-length neurofibromin is a GTPase activating protein (GAP) for the Ras proto-oncogene product. Regulation of neurofibromin activity therefore has important implications for cell growth. Neurofibromin co-purifies with tubulin when expressed in insect cells. The interaction between neurofibromin and tubulin is sensitive to the microtubule depolymerizing agent colchicine. Neurofibromin GAP activity is inhibited even at low concentrations of tubulin. However, maximal inhibition of GAP activity is only approximately 70%, suggesting that the neurofibromin-tubulin complex retains residual GAP activity. This decreased activity is reflected by a 4-fold decrease in its affinity for Ras. A truncated mutant of neurofibromin with reduced sensitivity to tubulin localizes some tubulin-binding determinants to an 80 residue segment immediately N-terminal to the GAP-related domain. Since tubulin is an abundant protein in eukaryotic cells, the tubulin-neurofibromin interaction may regulate the Ras signalling pathway.
Bollag
G
G
Department of Molecular Biology, Onyx Pharmaceuticals, Richmond, CA 94806.
McCormick
F
F
Clark
R
R
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001665:Binding Sites; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular; D020690:GTPase-Activating Proteins; D008969:Molecular Sequence Data; D009036:Moths; D025542:Neurofibromin 1; D011506:Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism*; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D025542:Neurofibromin 1; D011506:Proteins; D014404:Tubulin; D020703:ras GTPase-Activating Proteins