CHEAS ROLE IN REGULATING CHEMOTACTIC SIGNAL TRANSDUCTION

该建议侧重于CheAS在调节信号中的作用 细菌趋化性的转导。 有证据表明，Cheas 与蛋白激酶CheAL和一种调节 Cheal活动，Chew。 为了确定CheAS是否调节 CheAL，系留细胞测定将用于比较鞭毛 表达不同比例的CheAL与CheAS的细胞的旋转行为。 为了鉴定与CheAS物理相互作用的趋化蛋白， CheAS的显性突变等位基因将被分离并构象 这些等位基因的抑制基因。 探索CheAS在以下方面的作用： 调节磷酸盐转移，从空提取物的蛋白质复合物 表达CheAL、CheAS、CheW和转换器亚群的细胞将被 使用多克隆抗CheA抗体免疫沉淀，并测定 通过CheAL将磷酸盐从[γ-32 P] ATP转移至CheAL。 除了 有助于实现理解 信号转导途径相互关联，使得细胞引起适当的 对环境刺激的反应，预计这项研究将有助于 了解蛋白激酶的替代形式所起的调节作用 参与信号转导，并参与翻译和结构 同相重叠基因的蛋白质产物之间的关系。

This proposal focuses on the role of CheAS plays in regulating signal transduction in bacterial chemotaxis. Evidence suggests that CheAS interacts with the protein kinase, CheAL, and a protein that regulates CheAL activity, CheW. To determine whether CheAS regulates the activity of CheAL, tethered cell assays will be used to compare the flagellar rotational behavior of cells that express varying ratios of CheAL to CheAS. To identify chemotaxis proteins with which CheAS physically interacts, dominant mutant alleles of CheAS will be isolated and conformational suppressors of these alleles identified. To explore the role of CheAS in regulating phosphate transfer, protein complexes from extracts of null cells expressing subsets of CheAL, CheAS, CheW and transducers will be immunoprecipitated using polyclonal anti-CheA antibodies and assayed for phosphate transfer by CheAL from [gamma-32P] ATP to CheAL. In addition to contributing to the long-term goal of understanding how components of signal transduction pathways interrelate such that cells elicit proper responses to environmental stimuli, it is expected this study will help to understand the regulatory role played by alternate forms of protein kinases involved in signal transduction and to the translational and structural relationships between protein products of in-phase overlapping genes.