Park Yun Yeon, Nam Hyun-Ja, Do Mihyang, Lee Jae-Ho
Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon, South Korea.
Genomic instability Research Center, Ajou University School of Medicine, Suwon, South Korea.
Exp Mol Med. 2016 Aug 5;48(8):e250. doi: 10.1038/emm.2016.61.
RSK2, also known as RPS6KA3 (ribosomal protein S6 kinase, 90 kDa, polypeptide 3), is a downstream kinase of the mitogen-activated protein kinase (MAPK) pathway, which is important in regulating survival, transcription, growth and proliferation. However, its biological role in mitotic progression is not well understood. In this study, we examined the potential involvement of RSK2 in the regulation of mitotic progression. Interestingly, depletion of RSK2, but not RSK1, caused the accumulation of mitotic cells. Time-lapse analysis revealed that mitotic duration, particularly the duration for metaphase-to-anaphase transition was prolonged in RSK2-depleted cells, suggesting activation of spindle assembly checkpoint (SAC). Indeed, more BubR1 (Bub1-related kinase) was present on metaphase plate kinetochores in RSK2-depleted cells, and depletion of BubR1 abolished the mitotic accumulation caused by RSK2 depletion, confirming BubR1-dependent SAC activation. Along with the shortening of inter-kinetochore distance, these data suggested that weakening of the tension across sister kinetochores by RSK2 depletion led to the activation of SAC. To test this, we analyzed the RSK2 effects on the stability of kinetochore-microtubule interactions, and found that RSK2-depleted cells formed less kinetochore-microtubule fibers. Moreover, RSK2 depletion resulted in the decrease of basal level of microtubule as well as an irregular distribution of mitotic spindles, which might lead to observed several mitotic progression defects such as increase in unaligned chromosomes, defects in chromosome congression and a decrease in pole-to-pole distance in these cells. Taken together, our data reveal that RSK2 affects mitotic progression by regulating the distribution, basal level and the stability of mitotic spindles.
RSK2,也被称为RPS6KA3(核糖体蛋白S6激酶,90 kDa,多肽3),是丝裂原活化蛋白激酶(MAPK)通路的下游激酶,在调节细胞存活、转录、生长和增殖方面起着重要作用。然而,其在有丝分裂进程中的生物学作用尚未得到充分了解。在本研究中,我们检测了RSK2在调节有丝分裂进程中的潜在作用。有趣的是,RSK2的缺失而非RSK1的缺失导致有丝分裂细胞的积累。延时分析显示,在缺失RSK2的细胞中,有丝分裂持续时间,特别是从中期到后期转换的持续时间延长,这表明纺锤体组装检查点(SAC)被激活。实际上,在缺失RSK2的细胞中,中期板动粒上存在更多的BubR1(Bub1相关激酶),并且BubR1的缺失消除了由RSK2缺失引起的有丝分裂积累,证实了BubR1依赖性SAC激活。随着动粒间距离的缩短,这些数据表明,RSK2缺失导致姐妹动粒间张力减弱,从而激活了SAC。为了验证这一点,我们分析了RSK2对动粒-微管相互作用稳定性的影响,发现缺失RSK2的细胞形成的动粒-微管纤维较少。此外,RSK2的缺失导致微管基础水平降低以及有丝分裂纺锤体分布不规则,这可能导致这些细胞中出现一些有丝分裂进程缺陷,如未对齐染色体增加、染色体排列缺陷以及两极间距离减小。综上所述,我们的数据表明RSK2通过调节有丝分裂纺锤体的分布、基础水平和稳定性来影响有丝分裂进程。