Fassler Jan S, West Ann H
Department of Biology, University of Iowa, Iowa City, Iowa, USA.
Methods Enzymol. 2010;471:291-317. doi: 10.1016/S0076-6879(10)71016-8. Epub 2010 Mar 1.
The histidine kinase-based signal transduction pathway was first uncovered in bacteria and is a prominent form of regulation in prokaryotes. However, this type of signal transduction is not unique to prokaryotes; over the last decade two-component signal transduction pathways have been identified and characterized in diverse eukaryotes, from unicellular yeasts to multicellular land plants. A number of small but important differences have been noted in the architecture and function of eukaryotic pathways. Because of the powerful genetic approaches and facile molecular analysis associated with the yeast system, the SLN1 osmotic response pathway in Saccharomyces cerevisiae is particularly useful as a eukaryotic pathway model. This chapter provides an overview of genetic and biochemical methods that have been important in elucidating the stimulus-response events that underlie this pathway and in understanding the details of a eukaryotic His-Asp phosphorelay.
基于组氨酸激酶的信号转导途径最初是在细菌中发现的,是原核生物中一种重要的调节形式。然而,这种信号转导类型并非原核生物所特有;在过去十年中,已经在从单细胞酵母到多细胞陆地植物等多种真核生物中鉴定并表征了双组分信号转导途径。在真核生物途径的结构和功能方面已经注意到一些微小但重要的差异。由于酵母系统具有强大的遗传学方法和便捷的分子分析手段,酿酒酵母中的SLN1渗透反应途径作为真核生物途径模型特别有用。本章概述了一些遗传和生化方法,这些方法对于阐明该途径背后的刺激-反应事件以及理解真核生物组氨酸-天冬氨酸磷酰基传递的细节非常重要。