SUMMARY
Human pre-mRNA splicing requires the removal of introns with highly variable lengths, from tens to over a million nucleotides. Therefore, mechanisms of intron recognition and splicing are likely not universal. Recently, we reported that splicing in a subset of human short introns with truncated polypyrimidine tracts depends on RBM17 (SPF45), instead of the canonical splicing factor U2AF heterodimer. Here, we demonstrate that SAP30BP, a factor previously implicated in transcriptional control, is an essential splicing cofactor for RBM17. In vitro binding and NMR analyses demonstrate that a U2AF-homology motif (UHM) in RBM17 binds directly to a newly identified UHM-ligand motif (ULM) in SAP30BP. We show that this RBM17–SAP30BP interaction is required to specifically recruit RBM17 to phosphorylated SF3B1 (SF3b155), a U2 snRNP component in active spliceosomes. We propose a unique mechanism for splicing in a subset of short introns, in which SAP30BP guides RBM17 in the assembly of active spliceosomes.
In brief Fukumura et al. discover a general splicing mechanism in a subset of human short introns with truncated polypyrimidine tracts. This splicing reaction is mediated by intermediary RBM17–SAP30BP complex, instead of the known U2AF heterodimer. SAP30BP binding to RBM17 may support RBM17 association with active phosphorylated SF3B1 in U2 snRNP.
Highlights
RBM17 (SPF45) is a splicing factor required for a subset of human short introns
SAP30BP is an essential cofactor, which interacts with RBM17 via UHM–ULM binding
RBM17 forms a weak complex with SAP30BP before its binding with SF3B1 in U2 snRNP
RBM17–SAP30BP complex supports RBM17 to be recruited to active phosphorylated SF3B1
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
↵5 Lead contact
Other statistical analyses (Welch's t-test) were applied in Figures 1C, 5 and Supplementary Figure S1. The statistical analysis (Wilcoxon rank-sum test) was added in Figure 2B. Using confocal fluorescence microscopy, Supplementary FigureS5B was re-analyzed with higher resolution. Accordingly, these figures with the legends were replaced. Minor modifications and corrections were applied in the text and figures.
