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Real-time visualization of spliceosome assembly reveals basic principles of splice site selection

GSE275098 Homo sapiens Expression profiling by high throughput sequencing 4 samples Submitted 2026/02/01 Platform GPL30173
Summary
The spliceosome is a megadalton protein-RNA complex which removes introns from pre-mRNA, yet the dynamic early assembly steps have not been structurally resolved. Specifically, how the spliceosome selects the correct 3’ splice site (3’SS) amongst highly similar non-functional sites is not known. Here, we develop a kinetic model of splice site selection based on single-molecule U2AF heterodimer imaging in vitro and in vivo. The model successfully predicts alternative splicing patterns and indicates that 3’SS selection occurs while U2AF is in complex with the spliceosome, not during initial binding. This finding indicates the spliceosome operates in a ‘partial’ kinetic proofreading regime, catalyzed in part by the helicase DDX42, which increases selectivity to the underlying U2AF binding site while still allowing for efficient forward progression.
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Direct links to NCBI, no account and no request form: the whole study as GSE275098_RAW.tar, processed values as the series matrix, the supplementary file directory, and per-sample supplementary files for any of the 4 samples. Raw sequencing reads are also available from ENA.

Also filed as BioProject PRJNA1149302 and SRA study SRP527125. Searching any of these in the dataset finder brings you back here.

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