Dynein is a transient kinetochore component whose binding is regulated by microtubule attachment, not tension.

Dynein is a transient kinetochore component whose binding is regulated by microtubule attachment, not tension.
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DOI:
10.1083/jcb.151.4.739
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发表时间:
2000-11-13
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Nicklas RB
Nicklas RB
中科院分区:
其他
文献类型:
--
作者:
King JM;Hays TS;Nicklas RB

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细胞质动力蛋白是唯一已知的能够驱动染色体向纺锤体两极运动的动粒蛋白。在蝗虫精母细胞中,动力蛋白免疫荧光染色在前中期着丝粒处是明亮的,而在中期着丝粒处是暗淡的。我们已经确定,这些染色强度的差异反映了与动粒相关的动力蛋白量的差异。中期动粒恢复明亮的动力蛋白染色,如果他们脱离纺锤体微管显微操作,并保持分离10分钟。我们发现,这种增加动力蛋白染色是不是由回缩或解蔽的动力蛋白后脱离。因此,动力蛋白确实是精母细胞动粒的一种瞬时成分。我们进一步表明,微管附着,而不是张力,调节动力蛋白定位在着丝粒。动力蛋白结合对微管的存在极其敏感:少于正常数量一半的动粒微管导致大多数动粒动力蛋白的丢失。因此,在有丝分裂的早期,在动粒开始附着在微管上后不久,大量的动力蛋白就离开了动粒。因此,这种动力蛋白组分的可能功能仅限于染色体的初始附着和运动和/或在有丝分裂检查点中的作用。
Cytoplasmic dynein is the only known kinetochore protein capable of driving chromosome movement toward spindle poles. In grasshopper spermatocytes, dynein immunofluorescence staining is bright at prometaphase kinetochores and dimmer at metaphase kinetochores. We have determined that these differences in staining intensity reflect differences in amounts of dynein associated with the kinetochore. Metaphase kinetochores regain bright dynein staining if they are detached from spindle microtubules by micromanipulation and kept detached for 10 min. We show that this increase in dynein staining is not caused by the retraction or unmasking of dynein upon detachment. Thus, dynein genuinely is a transient component of spermatocyte kinetochores. We further show that microtubule attachment, not tension, regulates dynein localization at kinetochores. Dynein binding is extremely sensitive to the presence of microtubules: fewer than half the normal number of kinetochore microtubules leads to the loss of most kinetochoric dynein. As a result, the bulk of the dynein leaves the kinetochore very early in mitosis, soon after the kinetochores begin to attach to microtubules. The possible functions of this dynein fraction are therefore limited to the initial attachment and movement of chromosomes and/or to a role in the mitotic checkpoint.
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