Poster abstracts

Poster number 52 submitted by Eduardo Paredes

Investigation of Natural and Selected Nucleic Acid Enzymes that depend on a Cofactor

Eduardo Paredes (Carnegie Mellon University), Joy Maity (Carnegie Mellon University)

Abstract:
The glmS riboswitch occurs in all gram-positive bacteria exerting feedback control over production of glucosamine-6-phosphate (GlcN6P) that is used in cell-wall synthesis. Recent biochemical and structural analyses suggest that GlcN6P binds to the RNA and act s directly to catalyze RNA cleavage. Through analyses of the cleavage reaction with normal and hyperactivated 5'-S-phosphorothiolate substrates as well as differing cofactors, we are investigating the mechanistic role of the putative GlcN6P cofactor. In addition, to determine the effectiveness of a GlcN6P cofactor in catalysis of RNA cleavage we are using in vitro selection (SELEX) methods to identify GlcN6P-dependent DNAzymes. Once we obtain the optimal sequences that catalyze RNA cleavage, we plan to examine further the role of GlcN6P though use of hyperactivated substrates and compare the cleavage mechanism of the GlcN6P-dependent DNAzyme to that of the natural glmS ribozyme.

Keywords: SELEX, Ribozyme, 5-s-phosphorothiolate