High-throughput assays probing protein-RNA interactions of eukaryotic translation initiation factors |
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Authors: | Galicia-Vázquez Gabriela Lindqvist Lisa Wang Xiaofeng Harvey Isabelle Liu Jing Pelletier Jerry |
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Affiliation: | a Department of Biochemistry, McGill University, Montreal, Que., Canada H3G 1Y6 b McGill Cancer Center, McGill University, Montreal, Que., Canada H3G 1Y6 |
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Abstract: | Protein-RNA interactions are involved in all facets of RNA biology. The identification of small molecules that selectively block such bimolecular interactions could provide insight into previously unexplored steps of gene regulation. Such is the case for regulation of eukaryotic protein synthesis where interactions between messenger RNA (mRNA) and several eukaryotic initiation factors govern the recruitment of 40S ribosomes (and associated factors) to mRNA templates during the initiation phase. We have designed simple fluorescence polarization-based high-throughput screening assays that query the binding of several translation factors to RNA and found that the mixed inhibitor p-chloromercuribenzoate interferes with poly(A) binding protein-RNA interaction. |
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Keywords: | Chemical biology Translation Small-molecule screening HTS assay p-Chloromercuribenzoate |
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