Characterization of Cellular Proteins and RNA Interacting with Ribosomal Protein L22
Jonathan Tucci
Mentor: Ingrid Ruf
The purpose of this research was to explore the interactions of human ribosomal protein L22 with certain
cellular proteins and RNA. Two RNA molecules were of interest, EBER-1 (Epstein-Barr Encoded RNA-1)
and hTR (human telomerase RNA). We analyzed the effects of EBER-1 on hTR and the telomerase
holoenzyme and the localization of L22 by using the strong affinity of biotin for avidin. We constructed a
biotin acceptor peptide (BAP) tagged L22 protein that was biotinylated in the presence of a biotin ligase
(BirA). Biotinylated BAPL22 was detected or isolated by immobilized avidin or fluorescent or horseradish
peroxidase (HRP) conjugated streptavidin. We were also able to quantify telomerase activity within different
Akata Burkitt lymphoma cell lines and the effects of EBER-1 on this telomerase activity. Results gained from
these methods showed that the biotinylation of L22 is a valuable method in visualizing the cellular
localization of L22. Additionally, the presence of EBER-1 showed no discernable effect on the amount of
telomerase activity in Akata Burkitt lymphoma cells.