EB1 promotes microtubule dynamics by recruiting Sentin in Drosophila cells.
EB1 promotes microtubule dynamics by recruiting Sentin in Drosophila cells.
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DOI：
10.1083/jcb.201101108
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发表时间：
2011-06-13
期刊：
影响因子：
--
通讯作者：
Goshima G
中科院分区：
文献类型：
--
作者：
Li W;Miki T;Watanabe T;Kakeno M;Sugiyama I;Kaibuchi K;Goshima G
微管+端调节剂EB1将Sentin和可能的微管聚合酶带到微管+端，以促进微管动力学。高度保守的EB1家族蛋白结合到微管的生长端，招募多种货物蛋白，对动态微管的体内生成至关重要。然而，目前尚不清楚这些微管+端主调控因子如何促进微管动力学。在本文中，我们鉴定了一种新的EB1货运蛋白Sentin。与EB1缺失类似，黑腹果蝇S2细胞的感觉素缺失导致微管暂停增加，并导致纺锤体形成更短，而不会取代生长中的微管中的EB1。我们证明了Sentin与EB1的关联对于其正端定位和功能至关重要。此外，通过表达EBN-Sentin融合蛋白，EB1的c端货物结合区被Sentin取代，挽救了EB1表型。XMAP215微管聚合酶的同源物Msps（迷你纺锤体）末端积累的敲低。这些结果表明，EB1通过招募货物蛋白Sentin和微管聚合酶到微管尖端来促进动态微管行为。
The microtubule plus end regulator EB1 brings Sentin and possibly a microtubule polymerase to microtubule plus ends to promote microtubule dynamics. Highly conserved EB1 family proteins bind to the growing ends of microtubules, recruit multiple cargo proteins, and are critical for making dynamic microtubules in vivo. However, it is unclear how these master regulators of microtubule plus ends promote microtubule dynamics. In this paper, we identify a novel EB1 cargo protein, Sentin. Sentin depletion in Drosophila melanogaster S2 cells, similar to EB1 depletion, resulted in an increase in microtubule pausing and led to the formation of shorter spindles, without displacing EB1 from growing microtubules. We demonstrate that Sentin’s association with EB1 was critical for its plus end localization and function. Furthermore, the EB1 phenotype was rescued by expressing an EBN-Sentin fusion protein in which the C-terminal cargo-binding region of EB1 is replaced with Sentin. Knockdown of Sentin attenuated plus end accumulation of Msps (mini spindles), the orthologue of XMAP215 microtubule polymerase. These results indicate that EB1 promotes dynamic microtubule behavior by recruiting the cargo protein Sentin and possibly also a microtubule polymerase to the microtubule tip.
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