Poster abstracts

Poster number 36 submitted by Sean Frisbie

RNA Pseudoknot Structure in 16S Helix 18 Destabilizes the Binding and Enzymatic Activity of Ribosomal RNA Modification Enzyme RsuA

Sanjaya Abeysirigunawardena (Kent State University), Kumudie Jayalath (Kent State University), Sean Frisbie (Kent State University )

Abstract:
Ribosomes are ribonucleoprotein complexes that are involved in protein
biosynthesis in cells. Ribosomes are generated from the assembly of
three different ribosomal RNAs (rRNAs) and more than 50 ribosomal
proteins (r-proteins). Though the ribosome assembly is well studied, the
impact of post-transcriptional rRNA modification enzymes and their
respective modifications on ribosomal assembly is not completely
understood. In response to the lack of understanding of preferred
substrates and the structural prerequisites for the stable binding and
function of the rRNA modification enzymes, we study thermodynamics
of protein RsuA to rRNA. My study focused on understanding the impact
of pseudoknot structure of 16S helix 18 on the binding of RsuA. Several
rRNA mutants that are incapable of forming 16S helix 18 pseudoknot
was generated by using site-directed mutagenesis. FRET-based binding
assay are used to measure the thermodynamic stability of RsuA-rRNA
complexes. These findings will enable us to elucidate thermodynamically
preferred mechanism of RsuA binding to rRNA.

References:
https://www.mdpi.com/2218-273X/10/6/841

Keywords: Helix 18, 16s RNA, Enzymatic Activity