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First published online September 3, 2008
doi: 10.1242/jcs.030544

Journal of Cell Science 121, 3092-3098 (2008)
Published by The Company of Biologists 2008
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Research Article

The GDP-dependent Rab27a effector coronin 3 controls endocytosis of secretory membrane in insulin-secreting cell lines

Toshihide Kimura1, Yukiko Kaneko1, Shogo Yamada1, Hisamitsu Ishihara2, Takao Senda3, Akihiro Iwamatsu4 and Ichiro Niki1,*

1 Department of Pharmacology, Oita University Faculty of Medicine, 1-1 Idaigaoka, Hasama, Yufu, Oita 879-5593, Japan
2 Division of Molecular Metabolism and Diabetes, Tohoku University Graduate School of Medicine, 2-1 Seiryo, Aoba, Sendai, Miyagi 980-8575, Japan
3 Department of Anatomy I, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan
4 Protein Research Network, Inc., 1-13-5 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan

* Author for correspondence (e-mail: niki{at}med.oita-u.ac.jp)

Accepted 2 July 2008

Rab27a is involved in the control of membrane traffic, a crucial step in the regulated secretion. Typically, the guanosine triphosphate (GTP)-bound form has been considered to be active and, therefore, searching for proteins binding to the GTP-form has been attempted to look for their effectors. Here, we have identified the actin-bundling protein coronin 3 as a novel Rab27a effector that paradoxically bound guanosine diphosphate (GDP)-Rab27a in the pancreatic β-cell line MIN6. Coronin 3 directly bound GDP-Rab27a through its β-propeller structure. The most important insulin secretagogue glucose promptly shifted Rab27a from the GTP- to GDP-bound form. Knockdown of coronin 3 by RNAi resulted in the inhibition of phogrin (an insulin-granule-associated protein) internalization and the uptake of FM4-64 (a marker of endocytosis). Similar results were reproduced by disruption of the coronin-3–GDP-Rab27a interaction with the dominant-negative coronin 3, and coexpression of the GDP-Rab27a mutant rescued these changes. Taken together, our results indicate that interaction of GDP-Rab27a and coronin 3 is important in stimulus-endocytosis coupling, and that GTP- and GDP-Rab27a regulates insulin membrane recycling at the distinct stages.

Key words: Rab27a, Coronin 3, Endocytosis, Insulin, GDP-dependent effector


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