KINASES AND CONTROL OF CELL-TYPE SPECIALIZATION

激酶和细胞类型特化的控制

• 批准号: 3297103
• 负责人: BEVERLY ERREDE
• 金额: $ 13.49万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3297103
• 关键词: Escherichia coli; Saccharomyces; alleles; antibody; biological signal transduction; cell differentiation; cell growth regulation; cell type; enzyme substrate; gene expression; genes; genetic manipulation; laboratory rabbit; molecular genetics; nucleic acid hybridization; point mutation; protein engineering; protein kinase; protein sequence

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.

已知蛋白激酶在调节中起关键作用 级联对于控制动物细胞增殖和 专业化。 此类监管网络的医学相关性 考虑到转变的状态，最好是 细胞失去了分化表型的各个方面，并且 在外部或正常的监管限制中扩散。 蛋白质 激酶是被确定为“癌基因” 合作代理人负责与之相关的转型 各种癌症。 酵母，酿酒酵母是研究的模型系统 控制细胞类型专业化的分子机制。 我们的 目的是定义两种蛋白激酶的作用 和Stell基因产物，在细胞专业化中。 我们将 调查：（1）调节时间和/或细胞类型的因素 蛋白激酶活性的特异性（2）结构改变 在可以改变其控制，特异性和的蛋白质激酶中 功能和（3）对照电路中与之相互作用的组件 蛋白激酶作为底物或调节剂。 原则 在此处发现，应该适用于运行的途径 多细胞真核生物。 Ste Gene产品将以大肠杆菌的目的表示 激酶活性和酶学的纯化和评估。 大肠杆菌表达的材料也将用于引起 兔子中的抗血清。 特定的抗体将用作 确定Ste Gene产物细胞类型特异性的试剂 活动，修改或本地化。 体外诱变将 应用于在Ste Gene中创建结构性改变 可能阻止功能或以其他方式修改特异性的产品 体内功能。 使用IN的伪证实研究 体外产生的Ste突变等位基因将用于识别 交配调节途径的其他组件。