@article {24, title = {cTag-PAPERCLIP Reveals Alternative Polyadenylation Promotes Cell-Type Specific Protein Diversity and Shifts Araf Isoforms with Microglia Activation.}, journal = {Neuron}, volume = {95}, year = {2017}, month = {2017 Sep 13}, pages = {1334-1349.e5}, abstract = {

Alternative polyadenylation (APA) is increasingly recognized to regulate gene expression across different cell types, but obtaining APA maps from individual cell types typically requires prior purification, a stressful procedure that can itself alter cellular states. Here, we describe a new platform, cTag-PAPERCLIP,~that generates APA profiles from single cell populations in intact tissues; cTag-PAPERCLIP requires no tissue dissociation and preserves transcripts in native states. Applying cTag-PAPERCLIP to profile four major cell types in the mouse brain revealed common APA preferences between excitatory and inhibitory neurons distinct from astrocytes~and microglia, regulated in part by neuron-specific~RNA-binding proteins NOVA2 and PTBP2. We further~identified a role of APA in switching Araf protein isoforms during microglia activation, impacting production of downstream inflammatory cytokines. Our results demonstrate the broad applicability of cTag-PAPERCLIP and a previously undiscovered role of APA in contributing to protein diversity between different cell types and cellular states within the brain.

}, keywords = {Animals, Antigens, Neoplasm, Astrocytes, Brain, Cells, Cultured, Female, Humans, Male, Mice, Microglia, Nerve Tissue Proteins, Neuro-Oncological Ventral Antigen, Neurons, Organ Specificity, Polyadenylation, Polypyrimidine Tract-Binding Protein, Protein Isoforms, Protein Serine-Threonine Kinases, RNA-Binding Proteins}, issn = {1097-4199}, doi = {10.1016/j.neuron.2017.08.024}, author = {Hwang, Hun-Way and Saito, Yuhki and Park, Christopher Y and Blach{\`e}re, Nathalie E and Tajima, Yoko and Fak, John J and Zucker-Scharff, Ilana and Darnell, Robert B} }