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Published 13 March 2006. doi:10.1083/jcb.200511126
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 172, Number 6, 835-846
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Article

The selective continued linkage of centromeres from mitosis to interphase in the absence of mammalian separase



Kazuki Kumada1, Ryoji Yao1, Tokuichi Kawaguchi1, Mika Karasawa1, Yutaka Hoshikawa1, Koji Ichikawa1, Yoshinobu Sugitani1, Issei Imoto2, Johji Inazawa2, Minoru Sugawara3, Mitsuhiro Yanagida4, and Tetsuo Noda1,3

1 Japanese Foundation for Cancer Research, Cancer Institute, Tokyo 135-8550, Japan
2 Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113-8510, Japan
3 Center for Translational and Advanced Animal Research, Tohoku University School of Medicine, Sendai 980-8575, Japan
4 Department of Gene Mechanisms, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan

Correspondence to Tetsuo Noda: tnoda{at}jfcr.or.jp

Separase is an evolutionarily conserved protease that is essential for chromosome segregation and cleaves cohesin Scc1/Rad21, which joins the sister chromatids together. Although mammalian separase also functions in chromosome segregation, our understanding of this process in mammals is still incomplete. We generated separase knockout mice, reporting an essential function for mammalian separase. Separase-deficient mouse embryonic fibroblasts exhibited severely restrained increases in cell number, polyploid chromosomes, and amplified centrosomes. Chromosome spreads demonstrated that multiple chromosomes connected to a centromeric region. Live observation demonstrated that the chromosomes of separase-deficient cells condensed, but failed to segregate, although subsequent cytokinesis and chromosome decondensation proceeded normally. These results establish that mammalian separase is essential for the separation of centromeres, but not of the arm regions of chromosomes. Other cell cycle events, such as mitotic exit, DNA replication, and centrosome duplication appear to occur normally. We also demonstrated that heterozygous separase-deficient cells exhibited severely restrained increases in cell number with apparently normal mitosis in the absence of securin, which is an inhibitory partner of separase.

Abbreviations used in this paper: E, embryonic day; ES, embryonic stem; LSC, laser scanning cytometry; MEF, mouse embryonic fibroblast; MOI, multiplicity of infection; SKY, spectral karyotyping.


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