Poster abstracts

Poster number 9 submitted by Tupa Basu Roy

Dual binding specificity of 65kd RNA binding protein in the minor spliceosome

Tupa Basu Roy (BGES, Cleveland State University), Girish C Shukla (BGES, Cleveland State University)

Abstract:
Most eukaryotic nuclear-encoded genes are interrupted by introns, which are needed to be removed from pre-mRNAs by splicing to generate functional mRNAs. In metazoan, two types of introns are spliced by two distinct spliceosomes. The major type of introns or U2-dependent introns are spliced by U2-dependent spliceosomal small nuclear (sn) RNAs including U1, U2, U4, U5 and U6. On the other hand, minor or U12-dependent introns are spliced by a distinct set of snRNAs containing U11, U12, U4atac and U6atac snRNAs. U5 appears to be a common snRNA in both systems. Recent work shows that 3’ RNA element of U6atac snRNA is sufficient to guide U6 snRNA to minor spliceosome and activates the splicing of a U12-dependent intron. This data lead us to believe that splicing activity that targets U6atac to minor spliceosome is modulated by a RNA binding protein specific to 3’ stem-loop element of RNA element. Of various novel proteins of U12-dependent spliceosome, protein 65K 3’RRM contains a well–characterized domain called 3’ -RNA recognition motif and our preliminary data indicate that P65 3’ RRM protein has potential to bind U6atac 3’ stem-loop element. To determine 65K-C-RRM protein-RNA binding site on U6atac 3’ stem-loop, we are using Electrophoretic Mobility Shift Assay (EMSA) and a series of deletion mutants of U6atac RNAs. More investigations need to be done to determine dual binding specificity of 65kd RNA binding protein in the minor spliceosome.

Keywords: P65 3 RRM protein, U12-dependent spliceosome, U6atac 3 stem-loop element