Mallick Jaideep, Jansen Gregor, Wu Cunle, Whiteway Malcolm
Biology Department, Concordia University, 7141 Sherbrooke St. West, Montreal, QC, Canada, H4B 1R6.
Biochemistry Department, McGill University, 3655 Promenade Sir William Osler, Montreal, QC, Canada, H3G 1Y6.
Methods Mol Biol. 2016;1356:31-41. doi: 10.1007/978-1-4939-3052-4_3.
Many biological processes are regulated by protein-protein interactions, and the analysis of these interactions has been a productive endeavor contributing to our understanding of cellular organization and function. The yeast two-hybrid technique is a widely used, powerful method of analyzing protein-protein interactions. The currently used formats, however, have inherent limitations, providing an opportunity to develop new alternatives that extend our ability to detect protein-protein interactions of biological relevance. Here we present a two-hybrid system named SRYTH (Ste11p/Ste50p related yeast two-hybrid) based on the Ste11p/Ste50p interaction that uses the activation of the HOG pathway of Saccharomyces cerevisiae as a reporter for interactions. The system is suitable for detecting cytoplasmic protein interactions in their natural subcellular environment, and has been successfully used to investigate protein-protein interactions, including transcription factor associations, in Candida albicans.
许多生物过程都受蛋白质-蛋白质相互作用的调控，对这些相互作用的分析一直是一项卓有成效的工作，有助于我们理解细胞的组织和功能。酵母双杂交技术是一种广泛应用的、强大的分析蛋白质-蛋白质相互作用的方法。然而，目前使用的形式存在固有局限性，这为开发新的替代方法提供了机会，从而扩展我们检测具有生物学相关性的蛋白质-蛋白质相互作用的能力。在此，我们基于Ste11p/Ste50p相互作用提出了一种名为SRYTH（与Ste11p/Ste50p相关的酵母双杂交）的双杂交系统，该系统利用酿酒酵母的高渗甘油（HOG）途径的激活作为相互作用的报告基因。该系统适用于在其天然亚细胞环境中检测细胞质蛋白相互作用，并已成功用于研究白色念珠菌中的蛋白质-蛋白质相互作用，包括转录因子关联。
