Thiol-Containing RNA for the Study of Structure and Function of Ribozymes

Abstract

RNA performs multiple functions in cellular environments, such as transferring genetic information, catalyzing chemical reactions, and providing an integral component of ribonucleoprotein complexes involved in mRNA processing and translation. Many of these functions are poorly understood, mainly due to the lack of structural information. Because limited information has been obtained by physical and biophysical techniques, chemical and biochemical methods have been extensively used for studying RNA structure. This article outlines one such method which relies on site-specific incorporation of thiols into RNA. A brief overview of the methods for incorporation of thiols into RNA is followed by a detailed description of a procedure which utilizes postsynthetic modification of 2′-amino-containing RNA for incorporation of thiols. The use of thiol-containing RNA to form disulfide cross-links for the study of the structure and dynamics of ribozymes is subsequently described.