Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge, CB2 0RE, UK.
Centre for Cancer Cell and Molecular Biology, Barts Cancer Institute, Queen Mary University of London, London, EC1M 6BQ, UK.
Nat Commun. 2020 Apr 15;11(1):1851. doi: 10.1038/s41467-020-14978-7.
Genome stability relies on proper coordination of mitosis and cytokinesis, where dynamic microtubules capture and faithfully segregate chromosomes into daughter cells. With a high-content RNAi imaging screen targeting more than 2,000 human lncRNAs, we identify numerous lncRNAs involved in key steps of cell division such as chromosome segregation, mitotic duration and cytokinesis. Here, we provide evidence that the chromatin-associated lncRNA, linc00899, leads to robust mitotic delay upon its depletion in multiple cell types. We perform transcriptome analysis of linc00899-depleted cells and identify the neuronal microtubule-binding protein, TPPP/p25, as a target of linc00899. We further show that linc00899 binds TPPP/p25 and suppresses its transcription. In cells depleted of linc00899, upregulation of TPPP/p25 alters microtubule dynamics and delays mitosis. Overall, our comprehensive screen uncovers several lncRNAs involved in genome stability and reveals a lncRNA that controls microtubule behaviour with functional implications beyond cell division.
基因组稳定性依赖于有丝分裂和胞质分裂的适当协调，其中动态微管捕获并忠实分离染色体到子细胞中。通过针对超过 2000 个人类长非编码 RNA 的高内涵 RNAi 成像筛选，我们鉴定出许多参与细胞分裂关键步骤的长非编码 RNA，如染色体分离、有丝分裂持续时间和胞质分裂。在这里，我们提供的证据表明，染色质相关的长非编码 RNA linc00899 在多种细胞类型中耗尽时会导致强烈的有丝分裂延迟。我们对 linc00899 耗尽的细胞进行转录组分析，并鉴定出神经元微管结合蛋白 TPPP/p25 是 linc00899 的靶标。我们进一步表明，linc00899 结合 TPPP/p25 并抑制其转录。在耗尽 linc00899 的细胞中，TPPP/p25 的上调改变微管动力学并延迟有丝分裂。总的来说，我们的全面筛选揭示了几个参与基因组稳定性的长非编码 RNA，并揭示了一个长非编码 RNA 通过控制微管行为来影响细胞分裂以外的功能。
