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J. Biol. Chem., Vol. 278, Issue 22, 19826-19833, May 30, 2003

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Vps9p CUE Domain Ubiquitin Binding Is Required for Efficient Endocytic Protein Traffic^*

Brian A. Davies , Justin D. Topp , Agnel J. Sfeir , David J. Katzmann , Darren S. Carney , Gregory G. Tall ¶, Andrew S. Friedberg ||, Li Deng **, Zhijian Chen ** and Bruce F. Horazdovsky  

From the Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, ^¶Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, ^||Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390, ^**Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390

Rab5 GTPases are key regulators of protein trafficking through the early stages of the endocytic pathway. The yeast Rab5 ortholog Vps21p is activated by its guanine nucleotide exchange factor Vps9p. Here we show that Vps9p binds ubiquitin and that the CUE domain is necessary and sufficient for this interaction. Vps9p ubiquitin binding is required for efficient endocytosis of Ste3p but not for the delivery of the biosynthetic cargo carboxypeptidase Y to the vacuole. In addition, Vps9p is itself monoubiquitylated. Ubiquitylation is dependent on a functional CUE domain and Rsp5p, an E3 ligase that participates in cell surface receptor endocytosis. These findings define a new ubiquitin binding domain and implicate ubiquitin as a modulator of Vps9p function in the endocytic pathway.

Received for publication, January 31, 2003 , and in revised form, March 17, 2003.

^* This work was supported in part by grants from the National Institutes of Health (GM-055301) and by predoctoral fellowships from the Howard Hughes Medical Institute (to B. A. D.) and the National Science Foundation (to J. D. T.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 507-284-0308; Fax: 507-284-2053; E-mail: Horazdovsky.Bruce{at}mayo.edu.

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