Dataset for "Xist Repeat A coordinates an assembly of SR proteins to recruit SPEN and induce gene silencing"
Description
This dataset accompanies the manuscript entitled "Xist Repeat A coordinates an assembly of SR proteins to recruit SPEN and induce gene silencing." Xist represents a paradigm to understand mechanisms of RNA-mediated gene silencing in mammals, which remain unresolved. To induce silencing, Xist recruits the RNA-binding protein (RBP) SPEN through its Repeat A domain. Yet, how Repeat A recruits SPEN to enact silencing remains unclear. We report that Repeat A sequences critical for SPEN recruitment and Xist-induced silencing directly bind serine/arginine-rich (SR) proteins and not SPEN. One such protein, SRSF1, binds Repeat A with low-nanomolar affinity, binds SPEN via reciprocal RRM-RS interactions, and cooperates with SRSF2 to facilitate SPEN recruitment. SPEN and SR-protein-binding motifs promote Repeat A’s association with many proteins, including the m6A-methylation machinery and elongating RNA polymerase II. Furthermore, SPEN represses autosomal genes where its recruitment coincides with SR-protein binding. We reveal an essential role for SR proteins in coordinating Xist-induced silencing and suggest that the sensing of SR-protein-rich assemblies is an ancient mechanism by which SPEN targets genes for repression.
Files
Steps to reproduce
Several file names were corrupted upon upload to Mendeley Data. Due to space limitations in Mendeley Data, we were unable to upload raw microscopy image data. Requests for any data not included in this dataset will be fulfilled by the authors. Contact: jbt@unc.edu, jmcalabr@med.unc.edu.
Institutions
- University of North Carolina at Chapel HillNorth Carolina, Chapel Hill