KINASES AND CONTROL OF CELL-TYPE SPECIALIZATION

已知蛋白激酶在调节细胞凋亡中起关键作用。 级联反应对控制动物细胞增殖和 专业化 此类监管网络的医学相关性 最好通过考虑转换状态来理解，其中 细胞失去了其分化表型的某些方面， 在正常的监管限制之外扩散。 蛋白 激酶是已被鉴定为 协作代理，负责与 各种癌症。 酵母菌S.酿酒酵母是一个模型系统的研究 控制细胞类型特化的分子机制。 我们 目的是确定两种蛋白激酶的作用，STE 7 和STEII基因产物在细胞特化中的作用。 我们将 研究：（1）调节时间和/或细胞类型的因素 蛋白激酶活性特异性（2）结构改变 在蛋白激酶中，可以改变它们的控制、特异性和 功能和（3）控制电路中的组件， 蛋白激酶作为底物或作为调节剂。 原则 这里所揭示的应该适用于在 多细胞真核生物 STE基因产物将在E.大肠杆菌， 的纯化和激酶活性和酶学的评价。 急诊 大肠杆菌表达的材料也将用于引发 兔抗血清。 STE特异性抗体将用作 测定STE基因产物细胞类型特异性的试剂 活动、修改或定位。 体外诱变将 应用于在STE基因中产生结构改变 可能妨碍功能或改变特异性的产品 在体内的功能。 假回复突变研究 体外产生的STE突变等位基因将用于鉴定 交配调节途径的其他组成部分。

Protein kinases are known to play a pivotal role in regulatory cascades important to the control of animal cell proliferation and specialization. The medical relevance of such regulatory networks is best appreciated by considering the transformed state where cells have lost aspects of their differentiated phenotype and proliferate outside or normal regulatory constraints. Protein kinases are_among the "oncogenes" that have been identified as collaborative agents responsible for transformation associated with various cancers. The yeast, S. cerevisiae is a model system for the study of molecular mechanisms controlling cell-type specialization. Our objective is to define the role of two protein kinases, the STE7 and STEll gene products, in cell specialization. We will investigate: (1) factors that modulate the timing and/or cell-type specificity of protein kinase activity (2) structural alterations in protein kinases that can modify their control, specificity and function and (3) components in control circuits that interact with protein kinases as substrates or as regulators. The principles uncovered here should be applicable to pathways operating in multicellular eukaryotes. The STE gene products will be expressed in E. coli for the purpose of purification and evaluation of kinase activity and enzymology. The E. coli expressed material will also be used to elicit antisera in rabbits. The STE specific antibodies will be used as reagents to determine cell-type specificity of STE gene product activity, modification or localization. In vitro mutagenesis will be applied to create structural alterations in the STE gene products that may prevent function or otherwise modify specificity of function in vivo. Pseudoreversion studies utilizing the in vitro generated STE mutant alleles will be applied to identify other components of the mating regulatory pathway.