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© The Rockefeller University Press, 0021-9525/2000//1073 $5.00
The Journal of Cell Biology, Volume 149, Number 5, , 2000 1073-1086

Original Article

Zyxin, a Regulator of Actin Filament Assembly, Targets the Mitotic Apparatus by Interacting with H-Warts/Lats1 Tumor Suppressor



Toru Hirotaa,b, Tetsuro Morisakia, Yasuyuki Nishiyamaa, Tomotoshi Marumotoa, Kenji Tadac, Toshihiro Haraa, Norio Masukoa, Masaki Inagakid, Katsuyoshi Hatakeyamab, and Hideyuki Sayaa

a Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan
b 1st Department of Surgery, Niigata University School of Medicine, 1-757 Asahimachi-dori, Niigata 951-8510, Japan
c Department of Neurosurgery, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan
d Laboratory of Biochemistry, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya, Aichi 464-0021, Japan
Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan.+81-96-373-5120+81-96-373-5116

hsaya{at}gpo.kumamoto-u.ac.jp

The mitotic apparatus plays a pivotal role in dividing cells to ensure each daughter cell receives a full set of chromosomes and complement of cytoplasm during mitosis. A human homologue of the Drosophila warts tumor suppressor, h-warts/LATS1, is an evolutionarily conserved serine/threonine kinase and a dynamic component of the mitotic apparatus. We have identified an interaction of h-warts/LATS1 with zyxin, a regulator of actin filament assembly. Zyxin is a component of focal adhesion, however, during mitosis a fraction of cytoplasmic-dispersed zyxin becomes associated with h-warts/LATS1 on the mitotic apparatus. We found that zyxin is phosphorylated specifically during mitosis, most likely by Cdc2 kinase, and that the phosphorylation regulates association with h-warts/LATS1. Furthermore, microinjection of truncated h-warts/LATS1 protein, including the zyxin-binding portion, interfered with localization of zyxin to mitotic apparatus, and the duration of mitosis of these injected cells was significantly longer than that of control cells. These findings suggest that h-warts/LATS1 and zyxin play a crucial role in controlling mitosis progression by forming a regulatory complex on mitotic apparatus.

Key Words: Cdc2 • interaction • mitotic spindle • phosphorylation • serine/threonine kinase

© 2000 The Rockefeller University Press

Abbreviations used in this paper: DMPK, myotonic dystrophy protein kinase; GST, glutathione-S-transferase; HA, hemagglutinin 1; KLH, keyhole limpet-hemocyanin; X-gal, 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside.


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