REGULATION OF THE MITOSIS INHIBITING KINASE--WEEL

有丝分裂的正确时机是真核生物的关键要素 细胞周期。如果有丝分裂过早启动，那么对基因的损害 传递到子代细胞的物质可能发生。这样的基因损伤 在多细胞生物中是启动的一个促成因素 癌症的威胁。这个项目旨在定义一个关键的有丝分裂的调控。 抑制酶，Wee1。Wee1的调节将在#年研究 裂殖酵母，代表了一种优秀的模式 用于这类研究的真核系统。也就是说，监管要素 酵母菌的许多基本过程在人类身上保存得很好。一种激活剂 在有丝分裂中负责使Wee1失活的活动是 被指认出来。Cdc2被假定为代表这样的一种激酶。 然而，初步数据表明，在S.pombe中，cdc2不是主要的 此活动的贡献者。其他潜在的候选者，如Plo1 或者，CDS1将被测试为Wee1抑制激酶。如果这些是 排除，与N-末端相关的一个或多个激酶活性 Wee1将通过生物化学的方式进行纯化，因为它们与 GST-Wee1(N-末端)融合蛋白。一旦分离，该激酶(S)可以 快速测试它们在Wee1和细胞周期控制中的作用 酵母遗传学和生化技术。这些研究将提供 对有丝分裂的调控有更全面的了解，因此 致癌作用。

The correct timing of mitosis is a critical element of the eukaryotic cell cycle. If mitosis initiates prematurely then damage to the genetic material transmitted to daughter cells can occur. Such genetic damage in multicellular organisms is a contributory factor in the initiation of cancer. This project aims to define the regulation of a key mitotic inhibitory kinase, Wee1. The regulation of Wee1 will be studied in Schizzosaccharomyces pombe, which represents an excellent model eukaryotic system for such studies. That is, the regulatory elements of fundamental processes in yeast are well conserved in man. The kinase activity(ies) responsible for inactivating Wee1 at mitosis is (are) to be identified. Cdc2 has been postulated to represent such a kinase however, preliminary data suggest that in S.pombe, Cdc2 is not a major contributor to this activity. Other potential candidates such as Plo1 or Cds1 will be tested as Wee1 inhibitory kinases. If these are excluded, the kinase activity(ies) associated with the N-terminus of Wee1 will be purified biochemically, by virtue of their affinity for GST-Wee1(N-terminus) fusion proteins. Once isolated, the kinase(s) can be rapidly tested for their roles in Wee1 and cell cycle control using yeast genetics and biochemical techniques. These studies will provide a more complete understanding of the regulation of mitosis and therefore oncogenesis.