Investigating snRNA interactions of the GTPase Snu114p required for pre- messenger RNA splicing. (360G-Wellcome-079536_Z_06_Z)

The spliceosome is the RNA/protein complex that carries out pre-mRNA splicing.As splicing is certainly RNA catalysed, determining how proteins of the spliceosome interact with and control the functions of the snRNAs is an essential step in understanding splicing. One protein of the spliceosome, Snu114p, is a GTPase with homology to ribosomal tRNA translocase EF-2. Snu114p is proposed to be involved in U4/U6 snRNA unwinding, release of U1 andU4 snRNPs during spliceosome activation as well as repositioning of U5 snRNA between the two catalytic steps of splicing, similar to EF-2 tRNA repositioning. Little information, however, is available on how Snu114p actually interacts with these snRNAs in the spliceosome. Key goals are 1) produce mutants in Snu114p for tools to investigating how Snu114p interacts with the snRNAs during splicing. 2) Determine the snRNA sequence and structural requirements for interacting with Snu114p. 3) As EF-2 binds and is inhibited by the molecule sordarin, this molecule, and its derivatives, will be tested for binding to Snu114p and inhibition of splicing. This work will reveal how Snu114p interacts and controls the functions of essential RNA components of the spliceosome and may lead to development of a novel small molecule inhibitor of the spliceosome.