Mutational Analysis For The Further Characterization Of Saccharomyces cerevisiae Ribosomal Protein L30 And Its Interaction With The RPL30 Transcript

Cheryl Selah, Susan White

Department of Chemistry, Bryn Mawr College, Bryn Mawr, PA 19010

Abstract

The Saccharomyces cerevisiae ribosomal protein L30 is one of a unique group of proteins that bind to their own RNA in a feedback regulation mechanism. In order to study the interaction of L30 with its RPL30 transcript, two mutations have been made in L30 protein at a key position in the protein/RNA interface. Based on past NMR and current crystallographic studies and phylogenetic comparison of L30 proteins in other species, it is thought that aromatically conserved mutations at position 85 will maintain a native-like stabilizing interaction with a purine-rich bulge in the RNA and therefore, wild type binding efficiency. A phenylalanine at position 85 has been mutated to tryptophan and histidine, and the stability of the mutants in differing solvent conditions and their binding capabilities compared to the wild type protein have been investigated through circular dichroism, band shift, and filter binding experiments. The results of these assays are presented here.