Department of Experimental Oncology, European Institute of Oncology, I-20139 Milan, Italy.
J Cell Biol. 2010 Jul 12;190(1):73-87. doi: 10.1083/jcb.201001036.
The catalytic activity of the MPS1 kinase is crucial for the spindle assembly checkpoint and for chromosome biorientation on the mitotic spindle. We report that the small molecule reversine is a potent mitotic inhibitor of MPS1. Reversine inhibits the spindle assembly checkpoint in a dose-dependent manner. Its addition to mitotic HeLa cells causes the ejection of Mad1 and the ROD-ZWILCH-ZW10 complex, both of which are important for the spindle checkpoint, from unattached kinetochores. By using reversine, we also demonstrate that MPS1 is required for the correction of improper chromosome-microtubule attachments. We provide evidence that MPS1 acts downstream from the AURORA B kinase, another crucial component of the error correction pathway. Our experiments describe a very useful tool to interfere with MPS1 activity in human cells. They also shed light on the relationship between the error correction pathway and the spindle checkpoint and suggest that these processes are coregulated and are likely to share at least a subset of their catalytic machinery.
MPS1 激酶的催化活性对纺锤体组装检查点和有丝分裂纺锤体上染色体的双定向至关重要。我们报告说，小分子 reversine 是 MPS1 的一种有效的有丝分裂抑制剂。Reversine 以剂量依赖的方式抑制纺锤体组装检查点。将其添加到有丝分裂的 HeLa 细胞中会导致 Mad1 和 ROD-ZWILCH-ZW10 复合物从未附着的动粒中排出，这两者对于纺锤体检查点都很重要。通过使用 reversine，我们还证明 MPS1 是纠正不正确的染色体-微管附着所必需的。我们提供的证据表明，MPS1 作用于 AURORA B 激酶（错误修正途径的另一个关键成分）下游。我们的实验描述了一种非常有用的工具，可以干扰人细胞中的 MPS1 活性。它们还阐明了错误修正途径和纺锤体检查点之间的关系，并表明这些过程是共同调节的，并且可能至少共享其催化机制的一个子集。
