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Vol. 14, Issue 9, 3741-3752, September 2003

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Rabring7, a Novel Rab7 Target Protein with a RING Finger Motif

Kouichi Mizuno, Akiko Kitamura ^*, and Takuya Sasaki 

Department of Biochemistry, The University of Tokushima Graduate School of Medicine, Tokushima 770-8503, Japan

Submitted August 13, 2002; Revised April 7, 2003; Accepted May 23, 2003
Monitoring Editor: Suzanne Pfeffer

Rab7, a member of the Rab family small G proteins, has been shown to regulate intracellular vesicle traffic to late endosome/lysosome and lysosome biogenesis, but the exact roles of Rab7 are still undetermined. Accumulating evidence suggests that each Rab protein has multiple target proteins that function in the exocytic/endocytic pathway. We have isolated a new Rab7 target protein, Rabring7 (Rab7-interacting RING finger protein), using a CytoTrap system. It contains an H2 type RING finger motif at the C termini. Rabring7 shows no homology with RILP, which has been reported as another Rab7 target protein. GST pull-down and coimmunoprecipitation assays demonstrate that Rabring7 specifically binds the GTP-bound form of Rab7 at the N-terminal portion. Rabring7 is found mainly in the cytosol and is recruited efficiently to late endosomes/lysosomes by the GTP-bound form of Rab7 in BHK cells. Overexpression of Rabring7 not only affects epidermal growth factor degradation but also causes the perinuclear aggregation of lysosomes, in which the accumulation of the acidotropic probe LysoTracker is remarkably enhanced. These results suggest that Rabring7 plays crucial roles as a Rab7 target protein in vesicle traffic to late endosome/lysosome and lysosome biogenesis.

^* Present address: Department of Pediatrics, The University of Tokushima School of Medicine, Tokushima 770-8503, Japan. Abbreviations used: aa, amino acid; BMM, Burkholder's minimum media; BMM/glucose–leu, ura, glucose base BMM lacking leucine and uracil; BMM/galactose–leu, ura, galactose base BMM lacking leucine and uracil.

Corresponding author. E-mail address: sasaki{at}basic.med.tokushimau.ac.jp.

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