2013 Fiscal Year Final Research Report

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

Research Project

Project/Area Number	23790068
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Biological pharmacy
Research Institution	Hokkaido University
Principal Investigator	MAITA Hiroshi 北海道大学, 薬学研究科(研究院), 講師 (60431318)
Project Period (FY)	2011 – 2012
Keywords	分子生物学 / スプライシング / snRNP
Research Abstract	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.