摘要
微管骨架在细胞的有丝分裂、细胞迁移、神经分化和细胞内物质运输等生命重要过程起到不可或缺的作用。在细胞内,有效地对它的动态性进行调节对于完成这些功能至关重要。微管正端跟踪蛋白是一类特异定位到聚合微管正端的微管结合蛋白。它们能够调控微管的动态性,影响微管与细胞器和信号分子的作用,以及调节微管骨架网络所受到的力。它们对细胞形态和功能的多个方面都产生重要影响。为了更加深入的了解微管正端跟踪蛋白作用网络及其作用微管的机制,我利用信息学的方法鉴定了新的微管正端跟踪蛋白TIP150。它能够与微管末端蛋白EB1相互作用和共定位到聚合微管的末端。并且TIP150能与微管解聚酶MCAK作用,且相互作用受到Aurora B激酶的抑制。进一步的RNA干扰实验显示TIP150和MCAK的微管末端定位均依赖于EB1,且TIP150通过增强MCAK-EB1之间的相互作用发挥协助MCAK聚集到微管末端的功能。这些结果有助于我们进一步了解整个微管正端跟踪作用的动态性与可塑性。
同时,我的初步结果表明TIP150可能以四聚体的形式存在起到不同骨架间桥梁的作用。这暗示着TIP150在细胞迁移、纺锤体星状微管定位等细胞活动中潜在的生物学功能。
The microtubule(MT) cytoskeleton orchestrates cellular plasticity and dynamics underlying morphogenesis and cell division.Growing MT plus-ends have emerged as dynamic regulatory machinery in which specialized proteins,called plus-end-tracking proteins(+TIPs),bind and govern the plus-end dynamics essential for cell division and migration.However,the molecular mechanisms underlying the plus-end regulation by +TIPs at spindle and astral MT have remained elusive.Here we show that TIP150 is a novel +TIP.TIP150 binds to EB1 in vitro and co-localizes with EB1 at the MT plus-ends in vivo.Suppression of EB1 eliminates the plus-end-localization of TIP150.Interestingly,TIP150 also binds MCAK,a MT-depolymerase localizing to the plus-end of MT.Suppression of TIP150 diminishes the plus-end localization of MCAK.Importantly,Aurora B-mediated phosphorylation disrupts TIP150-MCAK association in vitro.I reason that TIP150 facilitates the EB1-dependent loading of MCAK onto MT plus-ends and orchestrates the dynamics at the plus-end of MTs.
My preliminary results also indicate that TIP150 is a tetramer and may play an important role in crosslinking different types of cytoskeleton,which suggest a possible function of TIP150 in the process of cell migration,astral microtubule position et al.
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