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Arrow(ARR)编码中心体蛋白(Cnn)。该基因具有多个组织特异性转录起始点，并可选择性地拼接产生不同形式蛋白质的mRNAs。基因的结构将通过绘制外显子、转录起始点和不同组织的cDNA序列来表征。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分析。以前的工作已经证明，CNN参与了果蝇的发育、配子发生和细胞生物学中的关键角色。对该蛋白可能功能的探讨(S)的重点将集中在它在中心体的无翼/Wnt(Wg)信号转导通路中的作用以及在卵母细胞的模式形成中的作用。一组CNN蛋白缺失结构将在培养的果蝇细胞中表达，以便在CNN中建立定位到中心体、响应Wg信号和被磷酸化所需的功能结构域。研究表明，CNN以一种细胞周期依赖的方式被磷酸化。这种磷酸化的参数将被确定。最终目标是确定CNN在WG信号通路中的位置，以确定其在发育中的不同角色的功能，并确定其在细胞骨架组织中的作用。信息是如何在细胞内以同步的方式传递的，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过程的洞察力。

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.