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Hiroshi Izuta^1,, Masashi Ikeno^2,, Nobutaka Suzuki^2,, Takeshi Tomonaga³, Naohito Nozaki⁴, Chikashi Obuse⁵, Yasutomo Kisu⁶, Naoki Goshima⁶, Fumio Nomura³, Nobuo Nomura⁶ and Kinya Yoda^1,*

¹ Bioscience and Biotechnology Center, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
² Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1101, Japan
³ Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan
⁴ Kanagawa Dental College, Inaoka-cho, Yokosuka-ku, Kanagawa 238-8580, Japan
⁵ Department of Gene Mechanism, Graduate School of Biostudies, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501, Japan
⁶ Biological Information Research Center, Advanced Industrial Science and Technology, Koto-ku, Tokyo135-0064, Japan

The centromere is a chromatin structure essential for correct segregation of sister chromatids, and defects in this region often lead to aneuploidy and cancer. We have previously reported purification of the interphase centromere complex (ICEN) from HeLa cells, and have demonstrated the presence of 40 proteins (ICEN1–40), along with CENP-A, -B, -C, -H and hMis6, by proteomic analysis. Here we report analysis of seven ICEN components with unknown function. Centromere localization of EGFP-tagged ICEN22, 24, 32, 33, 36, 37 and 39 was observed in transformant cells. Depletion of each of these proteins by short RNA interference produced abnormal metaphase cells carrying misaligned chromosomes and also produced cells containing aneuploid chromosomes, implying that these ICEN proteins take part in kinetochore functions. Interestingly, in the ICEN22, 32, 33, 37 or 39 siRNA-transfected cells, CENP-H and hMis6 signals disappeared from all the centromeres in abnormal mitotic cells containing misaligned chromosomes. These results suggest that the seven components of the ICEN complex are predominantly localized at the centromeres and are required for kinetochore function perhaps through or not through loading of CENP-H and hMis6 onto the centromere.

These authors contributed equally to this work

^* Correspondence: E-mail: i45156a{at}cc.nagoya-u.ac.jp

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