Comprehensive profiling of post-transcriptional regulatory activity

A dynamic network of proteins modulate the translation of messenger RNA (mRNA) to meet the changing needs of the cell. RNA-binding proteins (RBPs) are the key elements in this network that determine whether a given mRNA is translationally activated or repressed, localized to a specific region or compartment within the cell, or degraded. Motivated by the important role RBPs play in gene expression, several groups have worked to identify hundreds of yeast RBPs and their mRNA targets, and the RNA-binding domains (RBDs) of these proteins have been revealed on a proteome-wide scale. Though these studies have provided important evidence for the physical interaction of these RBPs with their cognate mRNAs, it has proven more challenging to discover their regulatory functions. To answer the outstanding question of how RBPs regulate mRNA translation, we have adapted the tethering assay into a broad and powerful survey for regulatory activity on a proteome-wide scale. Our data provide functional information with domain-level resolution for known regulatory proteins. We also identify regulatory effects from proteins with other primary functions in the cell or whose cellular roles have yet to be characterized. Our global functional analysis reveals missing links in translational control and allows a more comprehensive understanding of RNA biology.