Effects of mitotic and tubulin mutations on microtubule architecture in actively growing protoplasts of Aspergillus nidulans.
Effects of mitotic and tubulin mutations on microtubule architecture in actively growing protoplasts of Aspergillus nidulans.
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DOI：
10.1083/jcb.99.3.830
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发表时间：
1984-09
期刊：
影响因子：
--
通讯作者：
Morris NR
中科院分区：
文献类型：
--
作者：
Gambino J;Bergen LG;Morris NR
我们用免疫荧光显微镜表征微管（MT）结构在野生型和突变体原生质体构巢曲霉在间期和有丝分裂。由于在A的完整菌丝中通过免疫荧光观察MT在技术上很困难。在nidulans中，我们开发了一种在不干扰细胞生理学的条件下去除细胞壁的方法，如由所得原生质体以正常速率进行核分裂和细胞周期突变体表型在限制性温度下表达的事实所证明的。间期细胞表现出广泛的细胞质MT网络。在有丝分裂过程中，胞质MT大部分消失，出现核内纺锤体。我们以前已经表明，benA 33 β-微管蛋白突变导致有丝分裂纺锤体的超稳定，我们已经提出了额外的间接证据表明，tubA 1和tubA 4 α-微管蛋白突变不稳定纺锤体MT。在本文中，我们表明，benA 33突变增加了细胞质的MTs以及纺锤体MTs的稳定性和tubA 1和tubA 4突变不稳定的纺锤体和细胞质的MTs。
We used immunofluorescent microscopy to characterize microtubule (MT) architecture in wild-type and mutant protoplasts of Aspergillus nidulans at interphase and at mitosis. Because the visualization of MTs by immunofluorescence is technically difficult in intact hyphae of A. nidulans, we developed a method for removing the cell wall under conditions that do not perturb cell physiology, as evidenced by the fact that the resulting protoplasts undergo nuclear division at a normal rate and that cell cycle mutant phenotypes are expressed at restrictive temperature. Interphase cells exhibited an extensive network of cytoplasmic MTs. During mitosis the cytoplasmic MTs mostly disappeared and an intranuclear mitotic spindle appeared. We have previously shown that the benA 33 beta-tubulin mutation causes hyperstabilization of the mitotic spindle, and we have presented additional indirect evidence that suggested that the tubA1 and tubA4 alpha-tubulin mutations destabilize spindle MTs. In this paper, we show that the benA33 mutation increases the stability of cytoplasmic MTs as well as spindle MTs and that the tubA1 and tubA4 mutations destabilize both spindle and cytoplasmic MTs.