Poster abstracts
Poster number 106 submitted by Rohan Munoth
Ribosome pausing analysis in Cryptococcus neoformans
Rohan Munoth (Department of Biological Sciences,Carnegie Mellon University), Simon Chow, (Department of Biological Sciences,Carnegie Mellon University), Gemma May (Department of Biological Sciences,Carnegie Mellon University), Corey Knowles (Department of Microbiology and Immunology, University at Buffalo Medical school), John Panepinto (Department of Microbiology and Immunology, University at Buffalo Medical school), Joel McManus (Department of Biological Sciences,Carnegie Mellon University)
Abstract:
Cryptococcus neoformans is an opportunistic fungal pathogen that results in an estimated 625,000 deaths annually worldwide. Ribosome pausing during translation elongation is a critical regulatory mechanism influencing protein folding, targeting, and mRNA stability. Previous studies have identified that ribosome stalling can be induced by specific nascent peptides interacting with the ribosome exit tunnel, with factors such as peptide hydrophobicity, polarity, and charge contributing to this process. While it is well-established that certain amino acids, like proline, induce ribosome stalling in many eukaryotic species, the precise role of nascent peptide characteristics in influencing ribosome pausing in C. neoformans has not been determined. We used a novel analysis pipeline to investigate how nascent peptide properties affect ribosome pausing in C. neoformans. Consistent with prior findings, our results confirm that most pause sites occur when proline is in the ribosomal P site. In addition, our analysis revealed that the hydrophobicity and polarity of the nascent peptide increase significantly as it approaches the pause site, a trend that aligns with previous reports in other species. Furthermore, we observed that the charge at the ribosomal P-site is more positive in pause-site peptides compared to non-pause-site peptides, suggesting electrostatic interactions play a role in the stalling process. Our comparison with prior research corroborates the role of specific amino acids in pausing while highlighting additional peptide characteristics that may influence this phenomenon. In future work, we will apply our pipeline to analyze pausing in datasets from Saccharomyces cerevisiae and Candida albicans to determine the extent to which the trends observed in C. neoformans are conserved across species and to identify pause sites conserved across fungi.
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Keywords: Ribosome pausing, Cryptococcus neoformans