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Molecular and Cellular Biology, December 2005, p. 10315-10328, Vol. 25, No. 23
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.23.10315-10328.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Constitutive Centromere Component CENP-50 Is Required for Recovery from Spindle Damage{ddagger}

Yukinori Minoshima,1,{dagger} Tetsuya Hori,2,{dagger} Masahiro Okada,2 Hiroshi Kimura,3 Tokuko Haraguchi,4 Yasushi Hiraoka,4 Ying-Chun Bao,1 Toshiyuki Kawashima,1 Toshio Kitamura,1* and Tatsuo Fukagawa2*

The Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan,1 Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies, Mishima, Shizuoka 411-8540, Japan,2 Nuclear Function and Dynamics Unit, HMRO, Graduate School of Medicine, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan,3 CREST of JST, Kansai Advanced Research Center, NICT, Kobe 651-2492, Japan4

Received 15 July 2005/ Returned for modification 30 August 2005/ Accepted 13 September 2005

We identified CENP-50 as a novel kinetochore component. We found that CENP-50 is a constitutive component of the centromere that colocalizes with CENP-A and CENP-H throughout the cell cycle in vertebrate cells. To determine the precise role of CENP-50, we examined its role in centromere function by generating a loss-of-function mutant in the chicken DT40 cell line. The CENP-50 knockout was not lethal; however, the growth rate of cells with this mutation was slower than that of wild-type cells. We observed that the time for CENP-50-deficient cells to complete mitosis was longer than that for wild-type cells. Centromeric localization of CENP-50 was abolished in both CENP-H- and CENP-I-deficient cells. Coimmunoprecipitation experiments revealed that CENP-50 interacted with the CENP-H/CENP-I complex in chicken DT40 cells. We also observed severe mitotic defects in CENP-50-deficient cells with apparent premature sister chromatid separation when the mitotic checkpoint was activated, indicating that CENP-50 is required for recovery from spindle damage.

* Corresponding author. Mailing address for Tatsuo Fukagawa: National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan. Phone: 81-55-981-6792. Fax: 81-55-981-6742. E-mail: tfukagaw{at}lab.nig.ac.jp. Mailing address for Toshio Kitamura: The Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan. E-mail: kitamura{at}ims.u-tokyo.ac.jp.

{ddagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{dagger} These authors contributed equally to this work.

Molecular and Cellular Biology, December 2005, p. 10315-10328, Vol. 25, No. 23
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.23.10315-10328.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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