Abstract

The faithful and rapid translation of proteins from genetic information is an essential feature of the ribosome. The general process of tRNA selection is governed by the ability of the ribosome to select for the aminoacylated tRNA (aa-tRNA) that matches the codon in its A-site. The efficiency and accuracy of this selection depends on the ability of nucleotides to form proper hydrogen bonds. While much is known about how chemical alterations of tRNA and rRNA can impact the fidelity of translation, less is known about how similar changes to mRNA affect decoding. In this work, we describe several studies aimed at investigating the impact of mRNA structural alterations on translation. We examine the impact of alkylative damage of mRNA both in vitro and in vivo and show that the accumulation of these adducts increases ribosomal stalling and subsequent rescue by the trans-translation pathway in bacteria. Additionally, we characterize the base pairing preferences of the most common oxidative damage adduct of mRNA, 8-oxoguanosine, on the ribosome. Finally, we show that the kink structure adopted by the phosphodiester backbone of mRNA on the ribosome impacts translation under sub-optimal base pairing conditions. Together, this works offers novel insight into the importance of mRNA structure in maintaining the speed and fidelity of translation.

Committee Chair

Hani Zaher

Committee Members

Doug Chalker, Robert Kranz, Nima Mosammaparast, Dmitri Nusinow,

Comments

Permanent URL: https://doi.org/10.7936/5kns-m285

Degree

Doctor of Philosophy (PhD)

Author's Department

Biology & Biomedical Sciences (Plant & Microbial Biosciences)

Author's School

Graduate School of Arts and Sciences

Document Type

Dissertation

Date of Award

Summer 8-15-2019

Language

English (en)

Author's ORCID

http://orcid.org/0000-0003-4956-3739

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