Date of Award

1-1-2018

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Biological Sciences

Content Description

1 online resource (ix, 69 pages) : color illustrations.

Dissertation/Thesis Chair

Gabriele Fuchs

Committee Members

Marlene Belfort, Cara Pager

Keywords

IRES, IRES-mediated, RACK1, Translation, Virus, Ribosomes, Genetic translation, Messenger RNA, Hepatitis C virus, Eukaryotic cells

Subject Categories

Biochemistry | Biology | Molecular Biology

Abstract

Due to its sheer number of interacting partners, core ribosomal protein RACK1 is a key player in many cellular processes and has been shown to play a vital role of translation initiation of the Hepatitis C virus RNA. The HCV 5′ untranslated region contains an internal ribosome entry site. IRES-mediated translation is a process employed in eukaryotes by select viruses and some cellular mRNAs by which translation initiation bypasses the canonical mRNA cap-dependent pathway by means of an RNA secondary structure (the IRES). While cap-dependent translation requires the recruitment of a suite of initiation factors, IRES-mediated translation requires few to no factors. Analysis of dual luciferase assays confirm the necessity of RACK1 in IRES-mediated translation of HCV but also illuminates the role of RACK1 in the IRES-mediated translation of virus (PV) and encephalomyocarditis virus (EMCV). In contrast, depletion of RACK1 had no effect on translation of the intergenic IRES of cricket paralysis virus (CrPV). Secondly, we examined translation in poliovirus by means of a plaque assay and demonstrated that depletion of RACK1 results in reduced plaque size. Thirdly, RACK1 was also found to be involved in the translation of a number of proposed cellular IRESs.

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