Analysis of the assembly and functional defect of the spliceosome associated with Retinitis pigmentosa

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The spliceosome is a highly dynamic macromolecular ribonucleoprotein (RNP) machine that catalyzes pre-mRNA splicing by assembling U1, U2, U4, U5, and U6 small nuclear (sn)RNPs. To process large numbers of introns, synthesis and recycling of snRNPs must be maintained within an appropriate range to avoid their shortage, which would cause a severe disease such as Retinitis pigmentosa. However, the mechanism that maintains cellular snRNP levels is unknown. We constructed an expression library of a luciferase fragment fused to core components of snRNPs and used it to isolate PRPF6 and U5-40K that specifically reconstitute luciferase activity in the U5 snRNP complex. We revealed that the reporter detects the effects of small molecules on the levels of the U5 snRNP-reporter protein complex. Our assay will be useful to identify the small molecule that can control the snRNP level in an appropriate range.