RRM
2008 Rustbelt RNA Meeting
RRM
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Poster abstracts
Abstract:
The goal of this research is to identify the post-transcriptional and post-translational modifications of ribosomal subunits produced from the presence of the antibiotic, erythromycin. Mass spectrometry (MS) based methods are applied to examine ribosome assembly defects in this research.
Escherichia coli is being used as the initial model system.
Four strains of E. coli K-12 (wild type, N281, N282, SK5665) are grown in the presence and absence of erythromycin at 27oC and 37oC. Liquid chromatography with ultraviolet absorption (LC-UV) and mass spectrometry (LC-MS) are used to identify post-transcriptional modifications arising in ribosomal RNA (rRNA) found by comparing treated cells to untreated cells grown at the same temperature. The ribosomal proteins of erythromycin treated cells are then compared to ribosomal proteins of erythromycin absent cells grown at the same temperature using matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS).
Stalled intermediates of strain SK5665 grown with erythromycin at 27oC have been compared to cells grown without the drug at 27oC, with and without erythromycin at 37oC, as well as to K12 cells grown with and without the drug at both temperatures. Stalled intermediate precursors result from 50S assembly inhibition by erythromycin. Current data suggests that there may be a lack of proteins in this stalled intermediate which is owed to an inability of the proteins to bind to the 23S rRNA in the correct order. The inability for proteins to bind may be due to missing a post-transcriptional modification(s) in the 23S rRNA, leading to misfolding of the 23S rRNA.
References:
1. Champney, W. S. Chittum, H. S. Journal of Bacteriology 1994, 176, 6192-6198.
2. Champney, W. S. Current Topics in Medicinal Chemistry 2003, 3, 929-947.
3. Crain, P. Methods in Enzymology, 1993, 193, 782-791.
Keywords: Nucleosides, High Pressure Liquid Chromatography (HPLC), Mass Spectrometry
