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Originally published as MBC in Press, 10.1091/mbc.E04-08-0747 on September 29, 2004

Vol. 15, Issue 12, 5420-5430, December 2004

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Rab9 GTPase Regulates Late Endosome Size and Requires Effector Interaction for Its Stability

Ian G. Ganley, Kate Carroll, Lenka Bittova, and Suzanne Pfeffer *

Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305-5307

Submitted August 27, 2004; Revised September 15, 2004; Accepted September 16, 2004
Monitoring Editor: Jennifer Lippincott-Schwartz

Rab9 GTPase resides in a late endosome microdomain together with mannose 6-phosphate receptors (MPRs) and the tail-interacting protein of 47 kDa (TIP47). To explore the importance of Rab9 for microdomain establishment, we depleted the protein from cultured cells. Rab9 depletion decreased late endosome size and reduced the numbers of multilamellar and dense-tubule–containing late endosomes/lysosomes, but not multivesicular endosomes. The remaining late endosomes and lysosomes were more tightly clustered near the nucleus, implicating Rab9 in endosome localization. Cells displayed increased surface MPRs and lysosome-associated membrane protein 1. In addition, cells showed increased MPR synthesis in conjunction with MPR missorting to the lysosome. Surprisingly, Rab9 stability on late endosomes required interaction with TIP47. Rabs are thought of as independent, prenylated entities that reside either on membranes or in cytosol, bound to GDP dissociation inhibitor. These data show that Rab9 stability is strongly influenced by a specific effector interaction. Moreover, Rab9 and the proteins with which it interacts seem critical for the maintenance of specific late endocytic compartments and endosome/lysosome localization.


Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E04–08–0747. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E04–08–0747.

Abbreviations used: ESCRT, endosomal complex required for transport; GDI, GDP dissociation inhibitor; LAMP1, lysosome associated membrane protein 1; MPR, mannose 6-phosphate receptor; siRNA, small-interfering RNA; TIP47, tail-interacting protein of 47 kDa.

* Corresponding author. E-mail address: pfeffer{at}stanford.edu.




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