Sridharan Vinidhra, Azuma Yoshiaki
a Department of Molecular Biosciences , University of Kansas , Lawrence , KS , USA.
Cell Cycle. 2016 Aug 17;15(16):2135-2144. doi: 10.1080/15384101.2016.1191713. Epub 2016 May 26.
Polo-like kinase 1 (Plk1)-interacting checkpoint helicase (PICH) localizes at the centromere and is critical for proper chromosome segregation during mitosis. However, the precise molecular mechanism of PICH's centromeric localization and function at the centromere is not yet fully understood. Recently, using Xenopus egg extract assays, we showed that PICH is a promiscuous SUMO binding protein. To further determine the molecular consequence of PICH/SUMO interaction on PICH function, we identified 3 SUMO-interacting motifs (SIMs) on PICH and generated a SIM-deficient PICH mutant. Using the conditional expression of PICH in cells, we found distinct roles of PICH SIMs during mitosis. Although all SIMs are dispensable for PICH's localization on ultrafine anaphase DNA bridges, only SIM3 (third SIM, close to the C-terminus end of PICH) is critical for its centromeric localization. Intriguingly, the other 2 SIMs function in chromatin bridge prevention. With these results, we propose a novel SUMO-dependent regulation of PICH's function on mitotic centromeres.
与Polo样激酶1(Plk1)相互作用的检查点解旋酶(PICH)定位于着丝粒,对有丝分裂期间染色体的正确分离至关重要。然而,PICH着丝粒定位及其在着丝粒功能中的精确分子机制尚未完全阐明。最近,我们利用非洲爪蟾卵提取物分析表明,PICH是一种泛素样小分子修饰相关蛋白(SUMO)结合蛋白。为了进一步确定PICH/SUMO相互作用对PICH功能的分子影响,我们在PICH上鉴定出3个SUMO相互作用基序(SIMs),并构建了一个SIM缺陷型PICH突变体。利用PICH在细胞中的条件性表达,我们发现了PICH的SIMs在有丝分裂期间的不同作用。虽然所有SIMs对于PICH定位于后期超细DNA桥上并非必需,但只有SIM3(第三个SIM,靠近PICH的C末端)对其着丝粒定位至关重要。有趣的是,另外2个SIMs在防止染色质桥形成中发挥作用。基于这些结果,我们提出了一种新的SUMO依赖的PICH在有丝分裂着丝粒上功能的调控机制。