Tubulin isotypes optimize distinct spindle positioning mechanisms during yeast mitosis.
Tubulin isotypes optimize distinct spindle positioning mechanisms during yeast mitosis.
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DOI：
10.1083/jcb.202010155
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发表时间：
2021-12-06
期刊：
影响因子：
--
通讯作者：
Gupta ML
中科院分区：
文献类型：
--
作者：
Nsamba ET;Bera A;Costanzo M;Boone C;Gupta ML
Nsamba等人。使用基因替换和全基因组筛选来显示萌芽酵母中的α-微管蛋白亚型，差异地招募Dynein和Kar9途径的组件到微管，并优先优化这些纺锤体定位机制。这些结果强调了微管蛋白亚型是如何支持不同的微管功能的。微管是动态的细胞骨架细丝，在广泛的细胞过程中是必不可少的。它们由微管蛋白聚合而成，微管蛋白是α-和β-亚基的异源二聚体。大多数真核生物表达多种类型的α-和β-微管蛋白，然而它们在任何生物体中的功能相关性仍然很大程度上是未知的。芽期酵母中的两个α-微管蛋白亚型Tub1和Tub3被认为是功能互换的，但它们各自的功能还没有得到严格的研究。在这里，我们开发了在WT细胞中以与总微管蛋白相当的水平表达单一微管蛋白亚型的其他相同基因的酵母菌株。利用全基因组筛选，我们揭示了同型和两种主要有丝分裂纺锤体定位机制之间的独特相互作用。我们进一步利用这些细胞来证明Tub1和Tub3通过分别差异地招募依赖Dy1和Kar9的机制的关键组件来优化纺锤体定位。我们的结果为微管蛋白亚型如何使高度保守的微管在不同的细胞过程中发挥作用提供了新的机制见解。
Nsamba et al. use gene replacement and genome-wide screening to show the α-tubulin isotypes in budding yeast differentially recruit components of the Dynein and Kar9 pathways to microtubules and preferentially optimize these spindle positioning mechanisms. The results highlight how tubulin isotypes underlie diverse microtubule functions. Microtubules are dynamic cytoskeleton filaments that are essential for a wide range of cellular processes. They are polymerized from tubulin, a heterodimer of α- and β-subunits. Most eukaryotic organisms express multiple isotypes of α- and β-tubulin, yet their functional relevance in any organism remains largely obscure. The two α-tubulin isotypes in budding yeast, Tub1 and Tub3, are proposed to be functionally interchangeable, yet their individual functions have not been rigorously interrogated. Here, we develop otherwise isogenic yeast strains expressing single tubulin isotypes at levels comparable to total tubulin in WT cells. Using genome-wide screening, we uncover unique interactions between the isotypes and the two major mitotic spindle positioning mechanisms. We further exploit these cells to demonstrate that Tub1 and Tub3 optimize spindle positioning by differentially recruiting key components of the Dyn1- and Kar9-dependent mechanisms, respectively. Our results provide novel mechanistic insights into how tubulin isotypes allow highly conserved microtubules to function in diverse cellular processes.
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