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果蝇中母源表达的γ微管蛋白37CD在雌性减数分裂和胚胎有丝分裂中的需求不同。

Maternally expressed gamma Tub37CD in Drosophila is differentially required for female meiosis and embryonic mitosis.

作者信息

Wilson P G, Borisy G G

机构信息

Laboratory of Molecular Biology, University of Wisconsin, Madison 53705, USA.

出版信息

Dev Biol. 1998 Jul 15;199(2):273-90. doi: 10.1006/dbio.1998.8900.

Abstract

We report functional analysis of gamma Tub37CD, a maternally synthesized gamma-tubulin that is highly expressed during oogenesis and utilized at centrosomes in precellular embryos. Two gamma Tub37CD mutants contained missense mutations that altered residues conserved in all gamma-tubulins and alpha- and/or beta-tubulins. A third gamma Tub37CD missense mutant identified a conserved motif unique to gamma-tubulins. A fourth gamma Tub37CD mutant contained a nonsense mutation and the corresponding premature stop codon generated a protein null allele. Immunofluorescence analysis of laid eggs and activated oocytes derived from the mutants revealed microtubules and meiotic spindles that were close to normal even in the absence of gamma Tub37CD. Eggs lacking the maternal gamma-tubulin were arrested in meiosis, indicative of a deficiency in activation. Analysis of meiosis with in vitro activation techniques showed that the cortical microtubule cytoskeleton of mature wild-type eggs was reorganized upon activation and expressed as transient assembly of cortical asters, and this cortical reorganization was altered in gamma Tub37CD mutants. In precellular embryos of partial loss of function mutants, spindles were frequently abnormal and cell cycle progression was inhibited. Thus, gamma Tub37CD functions differentially in female meiosis and in the early embryo; while involved in oocyte activation, it is apparently not required or plays a subtle role in formation of the female meiotic spindle which is acentriolar, but is essential for assembly of a discrete bipolar mitotic spindle which is directed by centrosomes organized about centrioles.

摘要

我们报告了γTub37CD的功能分析,γTub37CD是一种由母体合成的γ微管蛋白,在卵子发生过程中高度表达,并在前细胞胚胎的中心体中发挥作用。两个γTub37CD突变体包含错义突变,这些突变改变了所有γ微管蛋白以及α和/或β微管蛋白中保守的残基。第三个γTub37CD错义突变体鉴定出了γ微管蛋白特有的保守基序。第四个γTub37CD突变体包含一个无义突变,相应的过早终止密码子产生了一个蛋白质无效等位基因。对来自这些突变体的产卵和活化卵母细胞进行免疫荧光分析发现,即使在没有γTub37CD的情况下,微管和减数分裂纺锤体也接近正常。缺乏母体γ微管蛋白的卵子在减数分裂中停滞,表明激活存在缺陷。用体外激活技术分析减数分裂表明,成熟野生型卵子的皮质微管细胞骨架在激活时会重新组织,并表现为皮质星状体的短暂组装,而这种皮质重组在γTub37CD突变体中发生了改变。在功能部分丧失的突变体的前细胞胚胎中,纺锤体经常异常,细胞周期进程受到抑制。因此,γTub37CD在雌性减数分裂和早期胚胎中发挥不同的功能;虽然它参与卵母细胞激活,但显然在无中心粒的雌性减数分裂纺锤体形成中不是必需的或只起微妙作用,但对于由围绕中心粒组织的中心体指导的离散双极有丝分裂纺锤体的组装至关重要。

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