Hemmerich Peter, Weidtkamp-Peters Stefanie, Hoischen Christian, Schmiedeberg Lars, Erliandri Indri, Diekmann Stephan
Leibniz Institute for Age Research, Fritz Lipmann Institute, 07745 Jena, Germany. phemmer@fl i-leibniz.de
J Cell Biol. 2008 Mar 24;180(6):1101-14. doi: 10.1083/jcb.200710052. Epub 2008 Mar 17.
To investigate the dynamics of centromere organization, we have assessed the exchange rates of inner centromere proteins (CENPs) by quantitative microscopy throughout the cell cycle in human cells. CENP-A and CENP-I are stable centromere components that are incorporated into centromeres via a "loading-only" mechanism in G1 and S phase, respectively. A subfraction of CENP-H also stays stably bound to centromeres. In contrast, CENP-B, CENP-C, and some CENP-H and hMis12 exhibit distinct and cell cycle-specific centromere binding stabilities, with residence times ranging from seconds to hours. CENP-C and CENP-H are immobilized at centromeres specifically during replication. In mitosis, all inner CENPs become completely immobilized. CENPs are highly mobile throughout bulk chromatin, which is consistent with a binding-diffusion behavior as the mechanism to scan for vacant high-affinity binding sites at centromeres. Our data reveal a wide range of cell cycle-specific assembly plasticity of the centromere that provides both stability through sustained binding of some components and flexibility through dynamic exchange of other components.
为了研究着丝粒组织的动态变化,我们通过定量显微镜在人类细胞的整个细胞周期中评估了着丝粒内部蛋白质(CENPs)的交换率。CENP-A和CENP-I是稳定的着丝粒成分,分别在G1期和S期通过“仅加载”机制整合到着丝粒中。CENP-H的一个亚组分也稳定地结合着丝粒。相比之下,CENP-B、CENP-C以及一些CENP-H和hMis12表现出不同的、细胞周期特异性的着丝粒结合稳定性,其驻留时间从几秒到几小时不等。CENP-C和CENP-H在复制期间特异性地固定在着丝粒上。在有丝分裂中,所有着丝粒内部的CENPs都完全固定。CENPs在整个染色质中具有高度流动性,这与作为在着丝粒处扫描空缺高亲和力结合位点机制的结合-扩散行为一致。我们的数据揭示了着丝粒在细胞周期特异性组装方面具有广泛的可塑性,既通过某些成分的持续结合提供稳定性,又通过其他成分的动态交换提供灵活性。