U5 snRNA-specific proteins studies, RNA-protein interaction, assembly of the spliceosomal Sm complex, and function implications of splicing in Cyanidioschyson merolae

Splicing is an interesting step in the processing of the precursor messenger RNA (pre-mRNA) that involves removal of the non-coding sequences (introns) and ligation of the coding sequences (exons). 50% of genetic diseases exert their effects through errors in splicing (López-Bigas et al. 2005). Therefore, a better understanding of this process can ease the development of cures for these diseases through genetic therapy. I proposed to investigate splicing in Cyanidioschyzon merolae that possesses a simpler spliceosome comprised of four snRNPs and 68 splicing proteins. I present successful expression of the Dib1 protein, and co-expression and purification of the Sm complex. I was able to prove formation of the ring-shaped Sm complex by electron microscopy analysis and binding of the complex to U2, U4, and U5 snRNAs. This work also initiated an investigation of splicing as a vital process for C. merolae by blockage of this mRNA maturation step with morpholino oligonucleotides.