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Loss of function of the?Drosophila?Ninein-related centrosomal protein Bsg25D causes mitotic defects and impairs embryonic development

机译:果蝇Ninin相关的中央体蛋白Bsg25D的功能丧失导致有丝分裂缺陷并损害胚胎发育

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The centrosome-associated proteins Ninein (Nin) and Ninein-like protein (Nlp) play significant roles in microtubule stability, nucleation and anchoring at the centrosome in mammalian cells. Here, we investigate?Blastoderm specific gene 25D?(Bsg25D), which encodes the only?Drosophila?protein that is closely related to Nin and Nlp. In early embryos, we find that?Bsg25D?mRNA and Bsg25D protein are closely associated with centrosomes and astral microtubules. We show that sequences within the coding region and 3′UTR of?Bsg25D?mRNAs are important for proper localization of this transcript in oogenesis and embryogenesis. Ectopic expression of eGFP-Bsg25D from an unlocalized mRNA disrupts microtubule polarity in mid-oogenesis and compromises the distribution of the axis polarity determinant Gurken. Using total internal reflection fluorescence microscopy, we show that an N-terminal fragment of Bsg25D can bind microtubules?in vitro?and can move along them, predominantly toward minus-ends. While flies homozygous for a?Bsg25D?null mutation are viable and fertile, 70% of embryos lacking maternal and zygotic Bsg25D do not hatch and exhibit chromosome segregation defects, as well as detachment of centrosomes from mitotic spindles. We conclude that Bsg25D is a centrosomal protein that, while dispensable for viability, nevertheless helps ensure the integrity of mitotic divisions in?Drosophila.
机译:与中心体相关的蛋白质Ninin(Nin)和Ninein-like蛋白(Nlp)在微管稳定性,成核和锚定在哺乳动物细胞中的中心体中起重要作用。在这里,我们研究“胚芽母细胞特异性基因25D”(Bsg25D),该基因编码唯一与Nin和Nlp密切相关的“果蝇”蛋白。在早期胚胎中,我们发现?Bsg25D?mRNA和Bsg25D蛋白与中心体和星状微管密切相关。我们显示,在Bsg25D?mRNAs的编码区域和3'UTR内的序列对于该转录本在卵子发生和胚胎发生中的正确定位很重要。 eGFP-Bsg25D从异位mRNA异位表达在产卵中期破坏微管极性,并损害轴极性决定簇Gurken的分布。使用全内反射荧光显微镜,我们发现Bsg25D的N端片段可以在体外结合微管,并且可以沿着微管移动,主要向负端移动。尽管纯合absg25Dnull突变的果蝇是可行和可育的,但缺乏母本和合子Bsg25D的胚胎中有70%不会孵化,并显示出染色体分离缺陷,以及中心体脱离有丝分裂纺锤体。我们得出的结论是,Bsg25D是一种中心体蛋白,尽管对于生存力而言是必不可少的,但它有助于确保果蝇中有丝分裂的完整性。

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