Characterization of spliceosome assembly in cyanidioschyzon merolae.

Pre-mRNA splicing is the removal of intervening sequences from pre-messenger RNA in a reaction catalyzed by the spliceosome, which contains five small nuclear RNAs (snRNAs) and more than 100 proteins. Assembly of the spliceosome occurs in a highly ordered manner, making the spliceosome a very complex and dynamic particle. The spliceosome has been studied in yeast and humans but a simpler system would simplify splicing studies. Cyanidioschyzon merolae (Cm) has been shown to have a simpler spliceosome. The goal of this study was to characterize the Cm spliceosome beginning with the question of how large it is. To measure the size of the Cm spliceosome I used glycerol gradient centrifugation and assembly gels to study the assembly pathways. Lastly an attempt was made to study the components of Cm spliceosome by developing an assay in yeast (Saccharomyces cerevisiae) where small molecule inhibitors were used to stall the spliceososme, which could then be purified and its composition studied. --Leaf ii.