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Benoit Bouvrette, Louis Philip; Wang, Xiaofeng; Cody, Neal A.L; Bergalet, Julie; Lefebvre, Fabio Alexis; Bovaird, Samantha; Blanchette, Mathieu; Lécuyer, Éric (Institut de Recherches Clinique de Montréal, Montréal, Canada; Département de Biochimie, Université de Montréal, Montréal, Canada; School of Computer Science, McGill University, Montréal, Canada)
Post-transcriptional regulation is often processed in distinct subcellular locations. Assessing the intracellular distribution of RNA binding proteins (RBPs) and RNA will yield insights as to their functions in gene regulation and expression. First, through a collection of antibodies recognizing >300 RBPs, we characterize their localization via immuno-labeling and co-labelling with 12 markers for specific substructures within both subnuclear and cytoplasmic bodies. These results are collected in the RBP Image Database, which exhibits >225,000 images of RBP-marker pair with controlled vocabulary describing their localization. Second, to gain global insights into RNA subcellular localization extent and correlation with their encoded protein, we biochemically fractionated cells and performed RNA deep sequencing and mass spectrometry on RNA and protein samples extracted from nuclear, cytosolic, membrane and insoluble fractions. By visualizing the RNA and proteins asymmetrical distribution within each fraction with 3-simplexes, we demonstrated the prevalence of RNA localization and revealed a gradient of correlations between groups of mRNAs and their encoded proteins.