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Volume 272, Number 42, Issue of October 17, 1997 pp. 26153-26158
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Interaction between Cdc42Hs and RhoGDI Is Mediated through the Rho Insert Region

(Received for publication, April 7, 1997, and in revised form, July 10, 1997)

From the Department of Pharmacology, Veterinary Medical Center, Cornell University, Ithaca, New York 14853

Members of the Rho subfamily of GTP-binding proteins contain a region of amino acid sequence (residues 122-134) that is absent from other Ras-like proteins and is termed the Rho insert region. To address the functional role of this domain, we have constructed a Cdc42Hs/Ras chimera in which loop 8 from Ha-Ras was substituted for the region in Cdc42Hs that contains the 13-amino acid insert region. Our data indicate that the insert region of Cdc42Hs is not essential for its interactions with various target/effector molecules or for interactions with the guanine nucleotide exchange factor, Dbl, or the Cdc42 GTPase-activating protein (GAP). However, the regulation of GDP dissociation and GTP hydrolysis on Cdc42Hs by the Rho GDP-dissociation inhibitor (GDI) is extremely sensitive to changes in the insert region, such that a Cdc42Hs/Ha-Ras chimera that lacks this insert is no longer susceptible to a GDI-induced inhibition of GDP dissociation and GTP hydrolysis. The insensitivity to GDI activity is not due to the inability of the GDI molecule to bind to the Cdc42Hs/Ha-Ras chimera, and in fact, the GDI is fully capable of stimulating the release of this chimera from membranes.

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