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酵母有丝分裂纺锤体的动态定位:微管马达蛋白和皮质决定因素的作用

Dynamic positioning of mitotic spindles in yeast: role of microtubule motors and cortical determinants.

作者信息

Yeh E, Yang C, Chin E, Maddox P, Salmon E D, Lew D J, Bloom K

机构信息

Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-3280, USA.

出版信息

Mol Biol Cell. 2000 Nov;11(11):3949-61. doi: 10.1091/mbc.11.11.3949.

Abstract

In the budding yeast Saccharomyces cerevisiae, movement of the mitotic spindle to a predetermined cleavage plane at the bud neck is essential for partitioning chromosomes into the mother and daughter cells. Astral microtubule dynamics are critical to the mechanism that ensures nuclear migration to the bud neck. The nucleus moves in the opposite direction of astral microtubule growth in the mother cell, apparently being "pushed" by microtubule contacts at the cortex. In contrast, microtubules growing toward the neck and within the bud promote nuclear movement in the same direction of microtubule growth, thus "pulling" the nucleus toward the bud neck. Failure of "pulling" is evident in cells lacking Bud6p, Bni1p, Kar9p, or the kinesin homolog, Kip3p. As a consequence, there is a loss of asymmetry in spindle pole body segregation into the bud. The cytoplasmic motor protein, dynein, is not required for nuclear movement to the neck; rather, it has been postulated to contribute to spindle elongation through the neck. In the absence of KAR9, dynein-dependent spindle oscillations are evident before anaphase onset, as are postanaphase dynein-dependent pulling forces that exceed the velocity of wild-type spindle elongation threefold. In addition, dynein-mediated forces on astral microtubules are sufficient to segregate a 2N chromosome set through the neck in the absence of spindle elongation, but cytoplasmic kinesins are not. These observations support a model in which spindle polarity determinants (BUD6, BNI1, KAR9) and cytoplasmic kinesin (KIP3) provide directional cues for spindle orientation to the bud while restraining the spindle to the neck. Cytoplasmic dynein is attenuated by these spindle polarity determinants and kinesin until anaphase onset, when dynein directs spindle elongation to distal points in the mother and bud.

摘要

在出芽酵母酿酒酵母中,有丝分裂纺锤体移动到芽颈处预先确定的分裂平面对于将染色体分配到母细胞和子细胞中至关重要。星状微管动力学对于确保细胞核迁移到芽颈的机制至关重要。细胞核在母细胞中朝着与星状微管生长相反的方向移动,显然是被皮质处的微管接触“推动”。相比之下,朝着颈部并在芽内生长的微管促进细胞核朝着微管生长的相同方向移动,从而将细胞核“拉”向芽颈。在缺乏Bud6p、Bni1p、Kar9p或驱动蛋白同源物Kip3p的细胞中,“拉动”失败很明显。因此,纺锤极体向芽中的分离失去了不对称性。细胞质运动蛋白动力蛋白对于细胞核向颈部的移动不是必需的;相反,据推测它通过颈部促进纺锤体伸长。在缺乏KAR9的情况下,在后期开始之前,依赖动力蛋白的纺锤体振荡很明显,后期后依赖动力蛋白的拉力也很明显,其超过野生型纺锤体伸长速度的三倍。此外,在没有纺锤体伸长的情况下,动力蛋白介导的对星状微管的力足以将一组2N染色体通过颈部进行分离,但细胞质驱动蛋白则不行。这些观察结果支持一个模型,其中纺锤体极性决定因素(BUD6、BNI1、KAR9)和细胞质驱动蛋白(KIP3)为纺锤体向芽的定向提供方向线索,同时将纺锤体限制在颈部。细胞质动力蛋白在后期开始之前被这些纺锤体极性决定因素和驱动蛋白减弱,后期开始时,动力蛋白将纺锤体伸长引导到母细胞和芽中的远端点。

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