MKKS Is a Centrosome-shuttling Protein Degraded by Disease-causing Mutations via CHIP-mediated Ubiquitination

• DOI: 10.1091/mbc.e07-07-0631
• Publication Date: 2007-12-20T01:35:02Z
• Authors: Shoshiro Hirayama; Yuji Yamazaki; Akira Kitamura; Yukako Oda; Daisuke Morito; Katsuya Okawa; Hiroshi Kimura; Douglas M. Cyr; Hiroshi Kubota; Kazuhiro Nagata
• Keywords: Centrosome; 0301 basic medicine; Ubiquitin-Protein Ligases; Glycine; Group II Chaperonins; 610; Glutamic Acid; 540; Microtubules; Cell Line; Mice; Protein Transport; 03 medical and health sciences; Cytosol; Solubility; Mutation; Animals; Humans; Mutant Proteins; Protein Structure, Quaternary; Bardet-Biedl Syndrome; Proteasome Inhibitors; Protein Processing, Post-Translational; Molecular Chaperones

McKusick–Kaufman syndrome (MKKS) is a recessively inherited human genetic disease characterized by several developmental anomalies. Mutations in the MKKS gene also cause Bardet–Biedl syndrome (BBS), a genetically heterogeneous disorder with pleiotropic symptoms. However, little is known about how MKKS mutations lead to disease. Here, we show that disease-causing mutants of MKKS are rapidly degraded via the ubiquitin–proteasome pathway in a manner dependent on HSC70 interacting protein (CHIP), a chaperone-dependent ubiquitin ligase. Although wild-type MKKS quickly shuttles between the centrosome and cytosol in living cells, the rapidly degraded mutants often fail to localize to the centrosome. Inhibition of proteasome functions causes MKKS mutants to form insoluble structures at the centrosome. CHIP and partner chaperones, including heat-shock protein (HSP)70/heat-shock cognate 70 and HSP90, strongly recognize MKKS mutants. Modest knockdown of CHIP by RNA interference moderately inhibited the degradation of MKKS mutants. These results indicate that the MKKS mutants have an abnormal conformation and that chaperone-dependent degradation mediated by CHIP is a key feature of MKKS/BBS diseases.