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J. Biol. Chem., Vol. 279, Issue 44, 46182-46190, October 29, 2004

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The Sac3 Homologue shd1 Is Involved in Mitotic Progression in Mammalian Cells^*

Sefat-e- Khuda, Mikoto Yoshida, Yan Xing, Tatsuya Shimasaki¶, Motohiro Takeya||, Kazuhiko Kuwahara**, and Nobuo Sakaguchi

From the Departments of Immunology and ^||Cell Pathology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1, Honjo, Kumamoto 860-8556, Japan, ^¶Division of Isotope Science, Center for Resource Analysis, Institute of Resource Development and Analysis, Kumamoto University, 2-2-1, Honjo, Kumamoto 860-0811, Japan, and ^**PRESTO and CREST, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan

Saccharomyces Sac3 required for actin assembly was shown to be involved in DNA replication. Here, we studied the function of a mammalian homologue SHD1 in cell cycle progression. SHD1 is localized on centrosomes at interphase and at spindle poles and mitotic spindles, similar to -tubulin, at M phase. RNA interference suppression of endogenous shd1 caused defects in centrosome duplication and spindle formation displaying cells with a single apparent centrosome and down-regulated Mad2 expression, generating increased micronuclei. Conversely, increased expression of SHD1 by DNA transfection with shd1-green fluorescent protein (gfp) vector for a fusion protein of SHD1 and GFP caused abnormalities in centrosome duplication displaying cells with multiple centrosomes and deregulated spindle assembly with up-regulated Mad2 expression until anaphase, generating polyploidy cells. These results demonstrated that shd1 is involved in cell cycle progression, in particular centrosome duplication and a spindle assembly checkpoint function.

Received for publication, May 13, 2004 , and in revised form, August 12, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AJ131957.

^* This work was supported by Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and by grants from Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Agency. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains Supplemental Figs. 1 and 2.

These authors equally contributed to this work.

To whom correspondence should be addressed. Tel.: 81-96-373-5134; Fax: 81-96-373-5138; E-mail: nobusaka{at}kaiju.medic.kumamoto-u.ac.jp.

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