Drosophila Katanin-60 Depolymerizes and Severs at Microtubule Defects
Drosophila Katanin-60 Depolymerizes and Severs at Microtubule Defects
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DOI：
10.1016/j.bpj.2011.03.062
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发表时间：
2011-05-18
影响因子：
3.4
通讯作者：
Ross, Jennifer L.
中科院分区：
文献类型：
--
作者：
Diaz-Valencia, Juan Daniel;Morelli, Margaret M.;Ross, Jennifer L.
微管（MT）的长度和位置在细胞中受到严格控制。调节MT动力学的MT相关蛋白的一个新家族是MT切断酶。在这项工作中，我们研究如何katanin（p60），被认为是第一个发现的切断酶，结合和切断MTs通过单分子全内反射荧光显微镜。我们发现切断活动取决于卡他宁浓度。我们还发现，katanin可以删除微管蛋白二聚体从终端的MT，出现去微管。引人注目的是，katanin定位和切断在GMPCPP-微管蛋白和GDP-微管蛋白的界面处，表明其靶向于原辅基移位缺陷。最后，我们观察到结合持续时间，流动性和寡聚化是ATP依赖性的。
Microtubule (MT) length and location is tightly controlled in cells. One novel family of MT-associated proteins that regulates MT dynamics is the MT-severing enzymes. In this work, we investigate how katanin (p60), believed to be the first discovered severing enzyme, binds and severs MTs via single molecule total internal reflection fluorescence microscopy. We find that severing activity depends on katanin concentration. We also find that katanin can remove tubulin dimers from the ends of MTs, appearing to depolymerize MTs. Strikingly, katanin localizes and severs at the interface of GMPCPP-tubulin and GDP-tubulin suggesting that it targets to protofilament-shift defects. Finally, we observe that binding duration, mobility, and oligomerization are ATP dependent.