Abstract
Little is known about how RNA binding proteins cooperate to control splicing, and how stress pathways reconfigure these assemblies to alter splice site selection. We have shown previously that SRSF10 plays an important role in the Bcl-x splicing response to DNA damage elicited by oxaliplatin in 293 cells. Here, RNA affinity assays using a portion of the Bcl-x transcript required for this response led to the recovery of the SRSF10-interacting protein 14-3-3ε and the Sam68-interacting protein hnRNP A1. Although SRSF10, 14-3-3ε, hnRNP A1/A2 and Sam68 do not make major contributions to the regulation of Bcl-x splicing under normal growth conditions, upon DNA damage they become important to activate the 5′ splice site of pro-apoptotic Bcl-xS. Our results indicate that DNA damage reconfigures the binding and activity of several regulatory RNA binding proteins on the Bcl-x pre-mRNA. Moreover, SRSF10, hnRNP A1/A2 and Sam68 collaborate to drive the DNA damage-induced splicing response of several transcripts that produce components implicated in apoptosis, cell-cycle control and DNA repair. Our study reveals how the circuitry of splicing factors is rewired to produce partnerships that coordinate alternative splicing across processes crucial for cell fate.
Highlights
Alternative splicing generates proteomic diversity that drives biological complexity
By extending our analysis to other transcripts associated with the DNA damage response (DDR), we report that the combinatorial contribution of hnRNP A1/A2, Sam[68] and SRSF10 to alternative splicing of DDR-relevant transcripts is broadly reconfigured after DNA damage, allowing coordinated splicing decisions to control the production of components that orchestrate the cellular response to DNA damage
We identified a RNA polymerase II (RNAPII) pausing site within SB1 (Reg23) that represses the use of the Bcl-xS, and is relieved by the elongation and splicing-related factor TCERG128
Summary
Alternative splicing generates proteomic diversity that drives biological complexity. Documenting the combinatorial contribution of several RBPs in the regulation of a specific splicing decision remains an understudied question in splicing control, and only a few model systems have been used to explore this complexity (e.g. Bcl-x, c-src, CD45 and Fas[3,4,5,6]) Another emerging but still poorly understood area concerns how the interactions between regulatory RBPs are restructured following physiological cues and environmental stresses such as DNA damage. By extending our analysis to other transcripts associated with the DDR, we report that the combinatorial contribution of hnRNP A1/A2, Sam[68] and SRSF10 to alternative splicing of DDR-relevant transcripts is broadly reconfigured after DNA damage, allowing coordinated splicing decisions to control the production of components that orchestrate the cellular response to DNA damage
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call