Poster abstracts
Poster number 100 submitted by Cesar Martinez
Mapping translation initiation sites using the bacterial toxin RelE
Cesar Martinez (Biological Chemistry, University of Michigan), Ross Kaufhold (Biological Chemistry, University of Michigan ), Dr. Jailson Brito Querido (Biological Chemistry, University of Michigan), Dr. Rachel Niederer (Biological Chemistry, University of Michigan)
Abstract:
Translational control of gene expression is critical for diverse cellular processes such as differentiation and stress tolerance. Translation regulation is achieved through a combination of cis and trans regulatory features within mRNA. However, the full extent of these features and their mechanisms of action remain poorly understood. To identify and delineate important regulatory elements our lab utilizes Direct Analysis of Ribosome Targeting (DART) (1) to measure ribosome recruitment to thousands of RNAs in parallel. We are interested in extending this method to instances where we expect changes in translation start site selection, such as RAN (repeat‐associated non‐AUG) translation. To gain insights into start site fidelity we have utilized the bacterial toxin, RelE. RelE cleaves ribosome‐bound mRNA in the ribosomal A site, leaving a record of precisely where the ribosome was bound. Addition of RelE dramatically reduces protein output from the in vitro translation assay, consistent with RelE actively cleaving RNA as expected. Future work will apply this method to investigate translation initiation start site selection in RAN translation substrates. This could potentially reveal how certain neurodegenerative diseases manifest in the human body.
References:
1 Niederer, R. O., Rojas‐Duran, M. F., Zinshteyn, B. & Gilbert, W. V. Direct analysis of ribosome targeting illuminates thousand‐fold regulation of translation initiation. Cell Syst 13, 256‐264 e253 (2022). https://doi.org:10.1016/j.cels.2021.12.002
Keywords: translation, initiation, RelE