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3205.pdf (21.89 MB)
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Mass Spectrometry Analysis of Methylated Ribosomal RNA
Author Info
Rohlfs, Rebecca L
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1368024445
Abstract Details
Year and Degree
2013, PhD, University of Cincinnati, Arts and Sciences: Chemistry.
Abstract
The goal of this dissertation is to identify and characterize post-transcriptional modifications occurring during ribosome biogenesis. The field of bio-analytical chemistry is advanced by this research by increasing our ability to characterize and understand the process of ribosomal ribonucleic acid (rRNA) modification. The role of many post-transcriptional modifications is still largely unknown and speculated; therefore, it is essential to study the chronology of post-transcriptional modifications during ribosome biogenesis. The focus of these studies is to use liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MSn) in the detection of rRNA modifications. The first study was to characterize post-transcriptional modifications within erythromycin-induced stalled ribosomal assembly intermediates. Liquid chromatography - mass spectrometry (LC-MS) was used to identify the post-transcriptional rRNA modifications present in improperly folded ribosome assembly subunits and compare these modifications to those found in properly assembled subunits. Limitations to this study led to the development of a site-specific assay for the characterization of rRNA modifications. A selected reaction-monitoring (SRM) assay was designed to detect and quantify site-specific post-transcriptional rRNA modifications. The advantage of this method is the ability to site-specifically characterize a modification that may occur multiple times within an rRNA transcript. Since some modifications occur at different times during ribosome biogenesis, it is important to quantify each modification at different stages of ribosome assembly. The SRM assay was applied to characterize post-transcriptional rRNA modifications treated with erythromycin and compare those modifications to post-transcriptional rRNA modifications of untreated cells. The work presented in this dissertation can be applied to other organisms for the construction of a timeline for rRNA modification during ribosome biogenesis. This work has progressed the field of rRNA modification and ribosome analysis by developing a new tool for the site-specific identification and characterization of rRNA modifications.
Committee
Patrick Limbach, Ph.D. (Committee Chair)
Bruce Ault, Ph.D. (Committee Member)
Joseph Caruso, Ph.D. (Committee Member)
Pages
149 p.
Subject Headings
Analytical Chemistry
Keywords
ribosomal RNA
;
liquid chromatography
;
mass spectrometry
;
SRM
;
MRM
;
RNA modification
;
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Citations
Rohlfs, R. L. (2013).
Mass Spectrometry Analysis of Methylated Ribosomal RNA
[Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1368024445
APA Style (7th edition)
Rohlfs, Rebecca.
Mass Spectrometry Analysis of Methylated Ribosomal RNA.
2013. University of Cincinnati, Doctoral dissertation.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1368024445.
MLA Style (8th edition)
Rohlfs, Rebecca. "Mass Spectrometry Analysis of Methylated Ribosomal RNA." Doctoral dissertation, University of Cincinnati, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1368024445
Chicago Manual of Style (17th edition)
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Document number:
ucin1368024445
Download Count:
286
Copyright Info
© 2013, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.