Investigating Fyv6’s role in splicing through RNA sequencing and selective protein depletion

Abstract

Splicing of pre-mRNAs is an essential process for regulation of eukaryotic gene expression. Splicing is carried out by the spliceosome, a highly dynamic complex. Mutations within the spliceosome are responsible for various diseases in humans. Studies have recently provided evidence for a novel splicing factor Fyv6 in Saccharomyces cerevisiae to be the homologous protein to human splicing factor FAM192A1

. Our publication on Fyv6 elucidated some of its function, however further research is necessary to fully understand Fyv6’s role within the spliceosome. Herein I describe experiments that would aid in further exploring this splicing factor through RNA sequencing and construction of yeast strains with an inducible degraded or sequestered Fyv6 protein. We outline initial RNA-seq experiment results and our efforts to tag Fyv6 in yeast for selective depletion. Through this work, we aim to further our understanding of the protein components that form the spliceosome, and its mechanism of gene regulation.