Clark I, Giniger E, Ruohola-Baker H, Jan L Y, Jan Y N
Howard Hughes Medical Institute, University of California at San Francisco 94143-0724.
Curr Biol. 1994 Apr 1;4(4):289-300. doi: 10.1016/s0960-9822(00)00068-3.
During oogenesis in Drosophila, determinants that will dictate abdomen and germline formation are localized to the 'polar plasm' in the posterior of the oocyte. Assembly of the polar plasm involves the sequential localization of several messenger RNAs and proteins to the posterior of the oocyte, beginning with the localization of oskar mRNA and Staufen protein during stages 8 and 9 of oogenesis. The mechanism by which these two early components accumulate at the posterior is not known. We have investigated whether directed transport along microtubules could be used to accomplish this localization.
We have made a fusion protein composed of the bacterial beta-galactosidase enzyme as a reporter, joined to part of the plus-end-directed microtubule motor, kinesin, and have found that the fusion protein transiently localizes to the posterior of the oocyte during stages 8 and 9 of oogenesis. Treatment with the microtubule-depolymerizing agent colchicine prevents both the localization of the fusion protein and the posterior transport of oskar mRNA and Staufen protein. Furthermore, the fusion protein localizes normally in oocytes mutant for either oskar and staufen, but not in other mutants in which oskar mRNA and Staufen protein are mislocalized.
Association with a plus-end-directed microtubule motor can promote posterior localization of a reporter protein during oogenesis. The genetic requirements for this localization and its sensitivity to colchicine, both of which are shared with the posterior transport of oskar mRNA and Staufen protein, suggest that similar mechanism may function in both processes.
在果蝇卵子发生过程中,决定腹部和生殖系形成的决定因子定位于卵母细胞后部的“极质”中。极质的组装涉及几种信使核糖核酸和蛋白质顺序定位于卵母细胞后部,始于卵子发生第8和9阶段时osk基因信使核糖核酸和Staufen蛋白的定位。这两种早期成分在后部积累的机制尚不清楚。我们研究了沿微管的定向运输是否可用于完成这种定位。
我们构建了一种融合蛋白,由细菌β-半乳糖苷酶作为报告基因,与正端定向微管马达驱动蛋白的一部分连接,发现该融合蛋白在卵子发生的第8和9阶段短暂定位于卵母细胞后部。用微管解聚剂秋水仙碱处理可阻止融合蛋白的定位以及osk基因信使核糖核酸和Staufen蛋白的向后运输。此外,融合蛋白在osk和staufen突变的卵母细胞中正常定位,但在osk基因信使核糖核酸和Staufen蛋白定位错误的其他突变体中则不然。
与正端定向微管马达驱动蛋白的结合可促进卵子发生过程中报告蛋白的向后定位。这种定位的遗传要求及其对秋水仙碱的敏感性,这两者与osk基因信使核糖核酸和Staufen蛋白的向后运输相同,表明类似机制可能在这两个过程中起作用。