50] 5S RNA structure and function

Abstract

Publisher Summary This chapter describes the use of some earlier and some more recently developed approaches which have employed for the investigation of the 5S RNA molecule. The studies have helped to add new and important facts regarding structure and also to understand more about the functional implications of 5S RNA for the ribosome and the mechanism of translation. This chapter is divided into biochemical approaches, including limited enzymatic digestion and chemical modification studies as well as cross-linking. The chapter also describes genetic approaches, where the, construction expression, and structural as well as functional characterization of 5S RNA mutants are reported. In vitro mutagenesis methods have made it possible to obtain a number of base change mutations in the gene for the ribosomal 5S RNA. Oligonucleotide- directed mutagenesis has yielded three point mutants. These transversions have been placed at crucial sites within the 5S RNA structure: G41 (G to C), A66 (A to C), and U103 (U to G). Random bisulfite catalyzed mutations within the gene for the 5S RNA have, on the other hand, provided a catalog of single, double, and multiple transition mutations.