Zeng C J Tracy, Lee Y-R Julie, Liu Bo
Department of Plant Biology, University of California, Davis, California 95616, USA.
Plant Cell. 2009 Apr;21(4):1129-40. doi: 10.1105/tpc.109.065953. Epub 2009 Apr 21.
Although cells of flowering plants lack a structurally defined microtubule-organizing center like the centrosome, organization of the spindles and phragmoplasts in mitosis is known to involve the evolutionarily conserved gamma-tubulin complex. We have investigated the function of Arabidopsis thaliana NEDD1, a WD40 repeat protein related to the animal NEDD1/GCP-WD protein, which interacts with the gamma-tubulin complex. The NEDD1 protein decorates spindle microtubules (MTs) preferentially toward spindle poles and phragmoplast MTs toward their minus ends. A T-DNA insertional allele of the single NEDD1 gene was isolated and maintained in heterozygous sporophytes, and NEDD1's function in cell division was analyzed in haploid microspores produced by the heterozygote. In approximately half of the dividing microspores exhibiting aberrant MT organization, spindles were no longer restricted to the cell periphery and became abnormally elongated. After mitosis, MTs aggregated between reforming nuclei but failed to appear in a bipolar configuration. Consequently, defective microspores did not form a continuous cell plate, and two identical nuclei were produced with no differentiation into generative and vegetative cells. Our results support the notion that the plant NEDD1 homolog plays a critical role in MT organization during mitosis, and its function is likely linked to that of the gamma-tubulin complex.
尽管开花植物的细胞缺乏像中心体那样结构明确的微管组织中心,但已知有丝分裂中纺锤体和成膜体的组织涉及进化上保守的γ-微管蛋白复合体。我们研究了拟南芥NEDD1的功能,它是一种与动物NEDD1/GCP-WD蛋白相关的WD40重复蛋白,能与γ-微管蛋白复合体相互作用。NEDD1蛋白优先向纺锤体极装饰纺锤体微管(MTs),并向其负端装饰成膜体MTs。分离出单个NEDD1基因的T-DNA插入等位基因,并在杂合子孢子体中维持,然后在杂合子产生的单倍体小孢子中分析NEDD1在细胞分裂中的功能。在大约一半表现出异常MT组织的分裂小孢子中,纺锤体不再局限于细胞周边,而是变得异常伸长。有丝分裂后,MTs在重新形成的细胞核之间聚集,但未能呈现双极构型。因此,有缺陷的小孢子没有形成连续的细胞板,而是产生了两个相同的细胞核,没有分化为生殖细胞和营养细胞。我们的结果支持这样一种观点,即植物NEDD1同源物在有丝分裂期间的MT组织中起关键作用,其功能可能与γ-微管蛋白复合体的功能相关。