Centrosomin Function in the Cell Cycle, Gametogenesis, and in the Wingless Signaling Pathway

9728694考夫曼 位于黑腹果蝇基因组中的复杂基因座arrow（arrow）已被证明编码蛋白质Centrosomin（Cnn）。该基因具有多个组织特异性转录起始位点和选择性剪接的mRNA，产生不同形式的蛋白质。该基因的结构将通过绘制外显子、转录起始位点和对不同组织的cDNA进行测序来表征。这些信息将导致建造一个比现在更有效的救援结构。同时，将继续进行扩大的突变体分析。以前的工作已经证明，Cnn参与果蝇的发育，配子发生和细胞生物学的关键作用。 对这种蛋白可能功能的研究重点将集中在其在中心体Wingless/Wnt（Wg）信号转导途径和卵母细胞模式中的作用。一组Cnn蛋白缺失构建体将在培养的果蝇细胞中表达，以在Cnn内建立定位于中心体所需的功能结构域，以响应Wg信号传导并被磷酸化。研究表明，Cnn以细胞周期依赖性方式磷酸化。将确定该磷酸化的参数。最终目标是将Cnn定位在Wg信号通路中，以确定其在发育中的各种作用中的功能，并确定其在细胞骨架组织中的作用。 信息如何在细胞内以同步的方式传递，以及它们如何导致复杂有机体的形成，在很大程度上仍然是未知的。这项工作将扩大我们对这一进程的了解。

9728694 Kaufman The complex locus, arrow (arr), which resides in the genome of Drosophila melanogaster has been demonstrated to encode the protein, Centrosomin (Cnn). The gene has multiple tissue-specific transcription start sites and alternatively spliced mRNAs that produce different forms of the protein. The gene's structure will be characterized by mapping exons, transcription start sites and the sequencing of cDNAs from different tissues. Such information will lead to the construction of a more effective rescue construct than is now available. Concurrently, an expanded mutant analysis will be continued. Previous work has demonstrated that Cnn is involved in critical roles in development, gametogenesis and cell biology in Drosophila. The focus of the inquiry into possible function(s) of this protein will center on its role in the Wingless/Wnt(Wg) signal transduction pathway in the centrosome and in the patterning of the oocyte. A set of Cnn protein deletion constructs will be expressed in cultured Drosophila cells in order to establish the functional domains within Cnn that are required for localization to the centrosome, to respond to Wg signaling and to be phosphorylated. Studies have demonstrated that Cnn is phosphorylated in a cell-cycle dependent manner. The parameters of that phosphorylation will be determined. The ultimate goals are to position Cnn within the Wg signaling pathway to determine its function in its various roles in development and to determine its role in the organization of the cytoskeleton. How messages are delivered in a synchronized manner within cells and how they lead to the formation of a complex organism is still largely unknown. This work will expand our insights into that process.