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首页> 外文期刊>Current Biology: CB >In vivo dynamics of the rough deal checkpoint protein during Drosophila mitosis
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In vivo dynamics of the rough deal checkpoint protein during Drosophila mitosis

机译：果蝇有丝分裂期间粗略检查点蛋白的体内动力学

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摘要

Rough Deal (Rod) and Zw10 are components of a complex required for the metazoan metaphase checkpoint [1, 2] and for recruitment of dynein/dynactin to the kinetochore [3]. The Rod complex, like most classical metaphase checkpoint components, forms part of the outer domain of unattached kinetochores. Here we analyze the dynamics of a GFP-Rod chimera in living syncytial Drosophila embryos. Uniquely among checkpoint proteins, GFP-Rod robustly streams from kinetochores along microtubules, from the time of chromosome attachment until anaphase onset. Prometaphase and metaphase kinetochores continuously recruit new Rod, thus feeding the current. Rod flux from kinetochores appears to require biorientation but not tension because it continues in the presence of taxol. As with Mad2, kinetochore- and spindle-associated Rod rapidly turns over with free cytosolic Rod, both during normal mitosis and after colchicine treatment, with a t(1/2) of 25-45 s. GFP-Rod coimmunoprecipitates with dynein/dynactin, and in the absence of microtubules both Rod and dynactin accumulate on kinetochores. Nevertheless, Rod and dynein/dynactin behavior are distinguishable. We propose that the Rod complex is a major component of the fibrous corona and that the recruitment of Rod during metaphase is required to replenish kinetochore dynein after checkpoint conditions have been satisfied but before anaphase onset.
机译：粗加工（Rod）和Zw10是后生中期检查点[1、2]和将动力蛋白/动力蛋白募集到动线粒[3]所需的复合物的组成部分。像大多数经典的中期检查点组件一样，Rod复合体形成了未连接的动植物的外部区域的一部分。在这里，我们分析了生活在合胞体果蝇胚胎中的GFP杆嵌合体的动力学。在检查点蛋白中，GFP-Rod从染色体的附着时间到后期开始，都从动植物沿着微管强劲地流过。前中期和中期动植物不断募集新的罗德，从而提供电流。运动动物的杆状通量似乎需要生物取向，但不需要张力，因为它在紫杉醇的存在下持续存在。与Mad2一样，在有丝分裂正常和秋水仙碱处理后，动摇杆和纺锤体相关的杆迅速用游离的胞质杆翻转，t（1/2）为25-45 s。 GFP-Rod与动力蛋白/动力蛋白共沉淀，在没有微管的情况下，动力蛋白和动力蛋白都堆积在动植物上。尽管如此，Rod和dynein / dynactin的行为还是可以区分的。我们认为，Rod复合物是纤维电晕的主要组成部分，并且在满足检查点条件之后但在后期发病之前，需要在中期募集Rod来补充动粒动力蛋白。

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