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Characterization of the Arabidopsis Augmin Complex Uncovers Its Critical Function in the Assembly of the Acentrosomal Spindle and Phragmoplast Microtubule Arrays

机译:拟南芥Augmin复合体的表征发现其关键功能在组装线粒体纺锤体和芦苇微管阵列。

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摘要

Plant cells assemble the bipolar spindle and phragmoplast microtubule () arrays in the absence of the centrosome structure. Our recent findings in Arabidopsis thaliana indicated that AUGMIN subunit3 (AUG3), a homolog of animal dim γ-tubulin 3, plays a critical role in γ-tubulin–dependent nucleation and amplification during mitosis. Here, we report the isolation of the entire plant augmin complex that contains eight subunits. Among them, AUG1 to AUG6 share low sequence similarity with their animal counterparts, but AUG7 and AUG8 share homology only with proteins of plant origin. Genetic analyses indicate that the AUG1, AUG2, AUG4, and AUG5 genes are essential, as stable mutations in these genes could only be transmitted to heterozygous plants. The sterile aug7-1 homozygous mutant in which AUG7 expression is significantly reduced exhibited pleiotropic phenotypes of seriously retarded vegetative and reproductive growth. The aug7-1 mutation caused delocalization of γ-tubulin in the mitotic spindle and phragmoplast. Consequently, spindles were abnormally elongated, and their poles failed to converge, as were splayed to discrete positions rendering deformed arrays. In addition, the mutant phragmoplasts often had disorganized bundles with uneven edges. We conclude that assembly of arrays during plant mitosis depends on the augmin complex, which includes two plant-specific subunits.
机译:在没有中心体结构的情况下,植物细胞组装了双极纺锤体和芦苇微管阵列。我们在拟南芥中的最新发现表明,AUGMIN亚基3(AUG3)是动物昏暗的γ-微管蛋白3的同系物,在有丝分裂期间γ-微管蛋白依赖性成核和扩增中起关键作用。在这里,我们报告了包含八个亚基的整个植物augmin复合物的分离。其中,AUG1至AUG6与其动物对应物具有低序列相似性,但是AUG7和AUG8仅与植物来源的蛋白具有同源性。遗传分析表明,AUG1,AUG2,AUG4和AUG5基因是必不可少的,因为这些基因中的稳定突变只能传递至杂合植物。 AUG7表达显着降低的无菌aug7-1纯合突变体表现出严重阻碍营养和生殖生长的多效性表型。 aug7-1突变引起γ-微管蛋白在有丝分裂纺锤体和成膜细胞中的脱位。结果,纺锤体异常伸长,其磁极未能收敛,并扩展到离散位置,从而使阵列变形。另外,突变的原生质体通常具有杂乱的束,边缘不均匀。我们得出的结论是,在植物有丝分裂过程中阵列的组装取决于包括两个植物特异性亚基的Augmin复合物。

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