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Vol. 17, Issue 6, 2780-2788, June 2006

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RINT-1 Regulates the Localization and Entry of ZW10 to the Syntaxin 18 Complex

School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan

Submitted October 24, 2005; Revised March 17, 2006; Accepted March 20, 2006
Monitoring Editor: Benjamin Glick

RINT-1 was first identified as a Rad50-interacting protein that participates in radiation-induced G₂/M checkpoint control. We have recently reported that RINT-1, together with the dynamitin-interacting protein ZW10 and others, is associated with syntaxin 18, an endoplasmic reticulum (ER)-localized SNARE involved in membrane trafficking between the ER and Golgi. To address the role of RINT-1 in membrane trafficking, we examined the effects of overexpression and knockdown of RINT-1 on Golgi morphology and protein transport from the ER. Overexpression of the N-terminal region of RINT-1, which is responsible for the interaction with ZW10, caused redistribution of ZW10. Concomitantly, ER-to-Golgi transport was blocked and the Golgi was dispersed. Knockdown of RINT-1 also disrupted membrane trafficking between the ER and Golgi. Notably, silencing of RINT-1 resulted in a reduction in the amount of ZW10 associated with syntaxin 18, concomitant with ZW10 redistribution. In contrast, no redistribution or release of RINT-1 from the syntaxin 18 complex was observed when ZW10 expression was reduced. These results taken together suggest that RINT-1 coordinates the localization and function of ZW10 by serving as a link between ZW10 and the SNARE complex comprising syntaxin 18.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

* These authors contributed equally to this work.

Address correspondence to: Mitsuo Tagaya ( tagaya{at}ls.toyaku.ac.jp)

Abbreviations used: aa, amino acid; BFA, brefeldin A; ER, endoplasmic reticulum; GFP, green fluorescent protein; GST, glutathione S-transferase; mAb, monoclonal antibody; Man II, mannosidase II; RNAi, RNA interference; siRNA, short interfering RNA; SNARE, SNAP receptor; VSVG, vesicular stomatitis virus-encoded glycoprotein.

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