Bardet-Biedl syndrome (BBS) is a pleiotropically genetic disorder, whose etiology is linked to cilia. Mutations in the Arf/Arl-family GTPase Arl6 have been recently shown to be responsible for BBS type 3. Here we show that BBS mutations alter the guanine nucleotide-binding properties of Arl6. Specifically, substitution of 31st Threonine to Arginine selectively abrogates the GTP-binding ability of Arl6 without affecting GDP binding/dissociating properties. Furthermore, all the BBS Mutations in Arl6 result in low expression of the mutant proteins, Which can be restored by the inhibition of the proteasome. These findings implicate that Arl6 Mutants are destabilized and eliminated by the proteasome in cells, probably due to the altered nucleotide-binding properties. (C) 2009 Elsevier Inc. All rights reserved.