Mitchell Sarah F
Department of Chemistry and Biochemistry, Loyola Marymount University, Los Angeles, CA, USA.
Methods Mol Biol. 2021;2209:235-249. doi: 10.1007/978-1-0716-0935-4_15.
mRNA binding proteins regulate gene expression by controlling the processing, localization, decay, and translation of messenger RNAs (mRNAs). To fully understand these mechanisms of posttranscriptional gene regulation, it is necessary to identify the complete set of mRNA binding proteins. In recent years, several assays have been developed to accomplish this goal in a wide variety of organisms. This work describes a method for the systematic identification of mRNA binding proteins in Saccharomyces cerevisiae. This method applies in vivo UV cross-linking, affinity pull-down of polyA(+) mRNAs, and analysis by mass spectrometry to identify proteins that directly bind to mRNAs.
mRNA结合蛋白通过控制信使核糖核酸（mRNA）的加工、定位、降解和翻译来调节基因表达。为了全面理解这些转录后基因调控机制，有必要鉴定出mRNA结合蛋白的完整集合。近年来，已经开发了几种检测方法来在多种生物体中实现这一目标。这项工作描述了一种在酿酒酵母中系统鉴定mRNA结合蛋白的方法。该方法应用体内紫外线交联、聚腺苷酸（polyA(+)）mRNA的亲和下拉以及质谱分析来鉴定直接与mRNA结合的蛋白质。
