Ribotoxins and their applications in probing the topographical structure of ribosomes

Abstract

Ribotoxins are a group of ribosome-inactivating proteins (RIPs) isolated mostly from plants. They inactivate ribosomes by a mechanism as RNA N-glycosidase that removes a specific adenine base from the highly conserved "S/R domain" in the largest ribosomal RNA. In this review, we introduce the major results from our laboratory in recent years on the study of the structure and function of RIPs and ribosomes: [1] Purification and characterization of the enzymatic mechanism of RIPs. Several new RIPs were purified and their RNA N-glycosidase and supercoil-dependent DNA endonuclease activities were studied. [2] The topographical structure of ribosomes. The relationship between the structure and function of ribosomes, especially of the "S/R domain" in rat 28S rRNA, were investigated by means of RIPs and other chemical probes. [3] The cytotoxicity of two RIPs to carcinoma cells. [4] Several new methods for studying RIPs and probing the structure of ribosomes were developed, i.e., radioassays for RNA N-glycosidase, glycoprotein detection by fluorescent labeling on SDS-polyacrylamide gels, and methods for small RNA sequencing.