Protein Phosphorylation Laboratory, Francis Crick Institute, 1 Midland Rd, London, NW1 1AT, UK.
Cancer Research UK, Manchester Institute, Alderley Park, SK10 4TG, UK.
Nat Commun. 2020 Mar 13;11(1):1396. doi: 10.1038/s41467-020-15163-6.
The Aurora B abscission checkpoint delays cytokinesis until resolution of DNA trapped in the cleavage furrow. This process involves PKCε phosphorylation of Aurora B S227. Assessing if this PKCε-Aurora B module provides a more widely exploited genome-protective control for the cell cycle, we show Aurora B phosphorylation at S227 by PKCε also occurs during mitosis. Expression of Aurora B S227A phenocopies inhibition of PKCε in by-passing the delay and resolution at anaphase entry that is associated with non-disjunction and catenation of sister chromatids. Implementation of this anaphase delay is reflected in PKCε activation following cell cycle dependent cleavage by caspase 7; knock-down of caspase 7 phenocopies PKCε loss, in a manner rescued by ectopically expressing/generating a free PKCε catalytic domain. Molecular dynamics indicates that Aurora B S227 phosphorylation induces conformational changes and this manifests in a profound switch in specificity towards S29 TopoIIα phosphorylation, a response necessary for catenation resolution during mitosis.
极光 B 脱离检查点延迟胞质分裂,直到解决陷入分裂沟的 DNA。这个过程涉及 PKCε 对 Aurora B S227 的磷酸化。评估这个 PKCε-Aurora B 模块是否为细胞周期提供了更广泛的基因组保护控制,我们发现 PKCε 也在有丝分裂期间使 Aurora B 的 S227 发生磷酸化。Aurora B S227A 的表达模拟了 PKCε 的抑制作用,绕过了与非分离和姐妹染色单体连接相关的后期进入的延迟和解决。这种后期延迟的实现反映在 caspase 7 依赖性细胞周期切割后 PKCε 的激活中;caspase 7 的敲低模拟了 PKCε 的缺失,以异位表达/生成游离 PKCε 催化结构域的方式得到挽救。分子动力学表明,极光 B S227 的磷酸化诱导构象变化,这表现在对 S29 TopoIIα 磷酸化的特异性上发生了深刻的转变,这是有丝分裂过程中连接体分辨率所必需的反应。