Liu Niankun, Lasko Paul
Department of Biology, McGill University, 3649 Promenade Sir William Osler, Montréal, Québec, Canada H3G 0B1.
G3 (Bethesda). 2015 Mar 31;5(6):1025-34. doi: 10.1534/g3.115.017517.
Embryonic patterning in Drosophila melanogaster is initially established through the activity of a number of maternally expressed genes that are expressed during oogenesis. mRNAs from some of these genes accumulate in the posterior pole plasm of the oocyte and early embryo and localize further into RNA islands, which are transient ring-like structures that form around the nuclei of future primordial germ cells (pole cells) at stage 3 of embryogenesis. As mRNAs from several genes with known functions in anterior-posterior patterning and/or germ cell specification accumulate in RNA islands, we hypothesized that some other mRNAs that localize in this manner might also function in these developmental processes. To test this, we investigated the developmental functions of 51 genes whose mRNAs accumulate in RNA islands by abrogating their activity in the female germline using RNA interference. This analysis revealed requirements for ttk, pbl, Hip14, eIF5, eIF4G, and CG9977 for progression through early oogenesis. We observed dorsal appendage defects in a proportion of eggs produced by females expressing double-stranded RNA targeting Mkrn1 or jvl, implicating these two genes in dorsal-ventral patterning. In addition, posterior patterning defects and a reduction in pole cell number were seen in the progeny of Mkrn1 females. Because the mammalian ortholog of Mkrn1 acts as an E3 ubiquitin ligase, these results suggest an additional link between protein ubiquitination and pole plasm activity.
黑腹果蝇的胚胎模式最初是通过许多在卵子发生过程中表达的母源基因的活性建立的。其中一些基因的mRNA在卵母细胞和早期胚胎的后极质中积累,并进一步定位到RNA岛中,RNA岛是在胚胎发生第3阶段围绕未来原始生殖细胞(极细胞)的细胞核形成的短暂环状结构。由于来自几个在前后模式形成和/或生殖细胞特化中具有已知功能的基因的mRNA在RNA岛中积累,我们推测以这种方式定位的其他一些mRNA可能也在这些发育过程中发挥作用。为了验证这一点,我们通过RNA干扰消除51个mRNA在RNA岛中积累的基因在雌性生殖系中的活性,来研究它们的发育功能。该分析揭示了ttk、pbl、Hip14、eIF5、eIF4G和CG9977在早期卵子发生进程中的需求。我们观察到,表达靶向Mkrn1或jvl的双链RNA的雌性产生的一部分卵子出现背侧附属物缺陷,这表明这两个基因参与背腹模式形成。此外,在Mkrn1雌性的后代中观察到后模式形成缺陷和极细胞数量减少。由于Mkrn1的哺乳动物直系同源物作为一种E3泛素连接酶,这些结果表明蛋白质泛素化与极质活性之间存在额外的联系。