Protein-RNA specificity by high-throughput principal component analysis of NMR spectra.

Katherine M Collins, Alain Oregioni, Laura E Robertson, Geoff Kelly, Andres Ramos

Nucleic acids research 2015 Mar 31

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Defining the RNA target selectivity of the proteins regulating mRNA metabolism is a key issue in RNA biology. Here we present a novel use of principal component analysis (PCA) to extract the RNA sequence preference of RNA binding proteins. We show that PCA can be used to compare the changes in the nuclear magnetic resonance (NMR) spectrum of a protein upon binding a set of quasi-degenerate RNAs and define the nucleobase specificity. We couple this application of PCA to an automated NMR spectra recording and processing protocol and obtain an unbiased and high-throughput NMR method for the analysis of nucleobase preference in protein-RNA interactions. We test the method on the RNA binding domains of three important regulators of RNA metabolism. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Citation

Katherine M Collins, Alain Oregioni, Laura E Robertson, Geoff Kelly, Andres Ramos. Protein-RNA specificity by high-throughput principal component analysis of NMR spectra. Nucleic acids research. 2015 Mar 31;43(6):e41