Impact of Cancer‐Associated Mutations in SF3B1 on Yeast pre‐mRNA Splicing

Abstract

In eukaryotes, a mRNA maturation step called splicing removes non‐coding intronic regions from pre‐mRNAs. Splicing is conducted by an enzyme called the spliceosome which uses a variety of protein and snRNA components in order to perform its catalytic function. One of these highly conserved components is the U2 snRNP protein SF3B1. In humans, mutation of SF3B1 can lead to improper regulation of alternative splicing. Many human diseases such as myelodysplastic syndromes (MDS), endometrial cancer (UCEC), bladder carcinoma (BLCA), and acute myeloid leukemia (LAML) have been linked to SF3B1 mutation and dysregulation of alternative splicing.The goal of this study is to introduce SF3B1 mutations observed in UCEC, BLCA, or LAML cancer patients into the homologous yeast protein (HSH155) to gain further insight into the consequences of these mutations on splicing. I have used site‐directed mutagenesis to incorporate these mutations into the HSH155 gene on plasmids and then shuffled these plasmids into a budding yeast strain in which the HSH155 gene was deleted from the genome. I then used temperature‐dependent growth assays and an in vivo splicing reporter to measure phenotypic responses due to these point mutations. These experiments will identify how cancer‐linked mutations change conserved, fundamental steps carried out by the spliceosome and how perturbation of these steps can led to changes in alternative splicing.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.