Keryer-Bibens Cecile, Barreau Carine, Osborne H Beverley
UMR 6061 Génétique et Développement, CNRS/Université de Rennes 1, IFR 140 Génétique Fonctionnelle, Agronomie et Santé, Faculté de Médecine, CS 34317, 2 avenue du Pr. Léon Bernard, 35043 Rennes cedex, France.
Biol Cell. 2008 Feb;100(2):125-38. doi: 10.1042/BC20070067.
Many steps in the control of gene expression are dependent on RNA-binding proteins, most of which are bi-functional, in as much as they both bind to RNA and interact with other protein partners in a functional complex. A powerful approach to study the functional properties of these proteins in vivo, independently of their RNA-binding ability, is to attach or tether them to specifically engineered reporter mRNAs whose fate can be easily followed. Two tethering systems have been mainly used in eukaryotic cells, namely the MS2 coat protein system and the lambda N-B box system. In this review, we firstly describe several studies in which these tethering systems have been used and provide an overview of these applications. We next describe the major features of these two systems, and, finally, we highlight a number of points that should be considered when designing experiments using this approach.
基因表达调控中的许多步骤都依赖于RNA结合蛋白，其中大多数具有双功能，因为它们既能与RNA结合，又能在功能复合物中与其他蛋白质伙伴相互作用。一种在体内研究这些蛋白质功能特性而不依赖其RNA结合能力的有效方法是将它们附着或拴系到经过特殊设计的报告mRNA上，其命运易于追踪。两种拴系系统主要用于真核细胞，即MS2外壳蛋白系统和λ N-B盒系统。在本综述中，我们首先描述了使用这些拴系系统的几项研究，并概述了这些应用。接下来，我们描述这两种系统的主要特征，最后，我们强调了在设计使用这种方法的实验时应考虑的一些要点。
