Poster abstracts

Poster number 149 submitted by Madison Turley

Ribonucleoprotein Mutational Profiling of Telomerase RNA

Madison Turley (Michigan State University), Basma Klump (Michigan State University), Jens Schmidt (Michigan State University), Chase Weidmann (University of Michigan)

Abstract:
Telomere shortening is an issue that results from loss of end replication of chromosomes during mitosis. Stem and cancer cells express a ribonucleoprotein called telomerase to combat telomere shortening. Human telomerase consists of telomerase RNA (TR), telomerase reverse transcriptase (TERT), a protein of interest TCAB1, and the H/ACA complex (dyskerin, NHP2, NOP10, and GAR1). The complete assembly and biogenesis of telomerase is still unknown. A technique to analyze bound proteins to RNA is needed to investigate this. Since its development, high-throughput RNA sequencing has preceded various other mutational profiling and sequencing techniques for RNA. One issue with these techniques is that the reverse transcriptase is terminated at regions with bound protein, making the sequencing unable to provide data on RNA-protein assembly. Dr. Chase Weidemann developed a ribonucleoprotein mutational profiling (RNP-MaP) approach that allowed more insight into the assembly of TR and its associated proteins. Our findings provided information on RNA-protein interactions, showing that H2A-H2B and dyskerin binding is independent of the presence of TCAB1. However, TERT typically needs TCAB1 to be bound before it can bind to TR. These findings allowed us to make conclusions about the assembly of telomerase with respect to TR.

Keywords: telomerase, mutational profiling, ribonucleoprotein