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© The Rockefeller University Press, 0021-9525/2000//505 $5.00
The Journal of Cell Biology, Volume 148, Number 3, , 2000 505-518

Original Article

Adenomatous Polyposis Coli (APC) Protein Moves along Microtubules and Concentrates at Their Growing Ends in Epithelial Cells



Yuko Mimori-Kiyosuea, Nobuyuki Shiinaa, and Shoichiro Tsukitaa,b

a Tsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
b Department of Cell Biology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8315, Japan
Department of Cell Biology, Kyoto University Faculty of Medicine, Konoe-Yoshida, Sakyo-ku, Kyoto 606-8315, Japan.81 75-753-466081 75-753-4372

htsukita{at}mfour.med.kyoto-u.ac.jp

Adenomatous polyposis coli (APC) tumor suppressor protein has been shown to be localized near the distal ends of microtubules (MTs) at the edges of migrating cells. We expressed green fluorescent protein (GFP)-fusion proteins with full-length and deletion mutants of Xenopus APC in Xenopus epithelial cells, and observed their dynamic behavior in live cells. During cell spreading and wound healing, GFP-tagged full-length APC was concentrated as granules at the tip regions of cellular extensions. At higher magnification, APC appeared to move along MTs and concentrate as granules at the growing plus ends. When MTs began to shorten, the APC granules dropped off from the MT ends. Immunoelectron microscopy revealed that fuzzy structures surrounding MTs were the ultrastructural counterparts for these GFP signals. The COOH-terminal region of APC was targeted to the growing MT ends without forming granular aggregates, and abruptly disappeared when MTs began to shorten. The APC lacking the COOH-terminal region formed granular aggregates that moved along MTs toward their plus ends in an ATP-dependent manner. These findings indicated that APC is a unique MT-associated protein that moves along selected MTs and concentrates at their growing plus ends through their multiple functional domains.

Key Words: adenomatous polyposis coli • green fluorescent protein • microtubules • microtubule-associated proteins • epithelial cells

© 2000 The Rockefeller University Press

The online version of this article contains supplemental material.

Abbreviations used in this paper: aa, amino acids; AMP-PNP, 5'-adenylylimidodiphosphate; APC, adenomatous polyposis coli; GFP, green fluorescent protein; MAP, microtubule-associated protein; MT, microtubule; pAb, polyclonal antibody.


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