Poster abstracts
Poster number 124 submitted by Stephanie Wang
Evaluating the hybridization of backbone branched and spliceosomal RNAs
Stephanie Wang (Carnegie Mellon University, Department of Chemistry), Stephanie Mack (Carnegie Mellon University, Department of Chemistry), Subha R. Das (Carnegie Mellon University, Department of Chemistry)
Abstract:
In the pre-messenger RNA splicing process, introns or non-coding regions of RNA are removed as lariats that form a loop with the 5'-end of the intron attached to an internal branch-point adenosine via a 2',5'- phosphodiester linkage. The highly conserved U6 and U2 small nuclear RNA (snRNA) found in spliceosomes are known to form duplexes along the branch point and recently observed in the yeast cryo-EM structures. We aim to study these interactions in vitro with synthetic backbone branched RNAs (bbRNAs) that can mimic the lariat intron sequences near the branch point. Here, we study the effects of bbRNAs and the effects of the 2',5'- phosphodiester bond on the thermal stability of RNA duplexes that would form with U2 and U6 snRNAs and variants through optical thermal melting measurements. These data will enable us to design RNA-based hybridization and splicing assays.
Keywords: backbone branched RNA, splicing, melting temperature