PRPF8-mediated dysregulation of hBrr2 helicase disrupts human spliceosome kinetics and 5´-splice-site selection causing tissue-specific defects.

Autor: Atkinson R; Biosciences Institute, Newcastle University, Newcastle, UK., Georgiou M; Biosciences Institute, Newcastle University, Newcastle, UK., Yang C; Biosciences Institute, Newcastle University, Newcastle, UK., Szymanska K; Leeds Institute of Medical Research, University of Leeds, Leeds, UK., Lahat A; Department of Biosciences, Durham University, Durham, UK., Vasconcelos EJR; Leeds Omics, University of Leeds, Leeds, UK., Ji Y; Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany.; Institute of Clinical Chemistry, University Medical Center Göttingen, Göttingen, Germany., Moya Molina M; Biosciences Institute, Newcastle University, Newcastle, UK.; Newcells Biotech, Newcastle, UK., Collin J; Biosciences Institute, Newcastle University, Newcastle, UK., Queen R; Biosciences Institute, Newcastle University, Newcastle, UK., Dorgau B; Biosciences Institute, Newcastle University, Newcastle, UK., Watson A; Biosciences Institute, Newcastle University, Newcastle, UK.; Newcells Biotech, Newcastle, UK., Kurzawa-Akanbi M; Biosciences Institute, Newcastle University, Newcastle, UK., Laws R; Electron Microscopy Research Services, Newcastle University, Newcastle, UK., Saxena A; Biosciences Institute, Newcastle University, Newcastle, UK., Shyan Beh C; Biosciences Institute, Newcastle University, Newcastle, UK., Siachisumo C; Biosciences Institute, Newcastle University, Newcastle, UK., Goertler F; Department of Informatics, University of Bergen, Bergen, Norway., Karwatka M; Leeds Institute of Medical Research, University of Leeds, Leeds, UK., Davey T; Electron Microscopy Research Services, Newcastle University, Newcastle, UK., Inglehearn CF; Leeds Institute of Medical Research, University of Leeds, Leeds, UK., McKibbin M; Leeds Institute of Medical Research, University of Leeds, Leeds, UK., Lührmann R; Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany., Steel DH; Biosciences Institute, Newcastle University, Newcastle, UK., Elliott DJ; Biosciences Institute, Newcastle University, Newcastle, UK., Armstrong L; Biosciences Institute, Newcastle University, Newcastle, UK., Urlaub H; Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany.; Institute of Clinical Chemistry, University Medical Center Göttingen, Göttingen, Germany.; Göttingen Center for Molecular Biosciences, Georg August University of Göttingen, Göttingen, Germany., Ali RR; Centre for Cell and Gene Therapy, Kings College London, London, UK., Grellscheid SN; Department of Biosciences, Durham University, Durham, UK.; Department of Informatics, University of Bergen, Bergen, Norway., Johnson CA; Leeds Institute of Medical Research, University of Leeds, Leeds, UK. c.johnson@leeds.ac.uk., Mozaffari-Jovin S; Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany. sina.mozaffari-jovin@mpinat.mpg.de.; Department of Medical Genetics and Medical Genetics Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. sina.mozaffari-jovin@mpinat.mpg.de., Lako M; Biosciences Institute, Newcastle University, Newcastle, UK. majlinda.lako@ncl.ac.uk.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Apr 11; Vol. 15 (1), pp. 3138. Date of Electronic Publication: 2024 Apr 11.
DOI: 10.1038/s41467-024-47253-0
Abstrakt: The carboxy-terminus of the spliceosomal protein PRPF8, which regulates the RNA helicase Brr2, is a hotspot for mutations causing retinitis pigmentosa-type 13, with unclear role in human splicing and tissue-specificity mechanism. We used patient induced pluripotent stem cells-derived cells, carrying the heterozygous PRPF8 c.6926 A > C (p.H2309P) mutation to demonstrate retinal-specific endophenotypes comprising photoreceptor loss, apical-basal polarity and ciliary defects. Comprehensive molecular, transcriptomic, and proteomic analyses revealed a role of the PRPF8/Brr2 regulation in 5'-splice site (5'SS) selection by spliceosomes, for which disruption impaired alternative splicing and weak/suboptimal 5'SS selection, and enhanced cryptic splicing, predominantly in ciliary and retinal-specific transcripts. Altered splicing efficiency, nuclear speckles organisation, and PRPF8 interaction with U6 snRNA, caused accumulation of active spliceosomes and poly(A)+ mRNAs in unique splicing clusters located at the nuclear periphery of photoreceptors. Collectively these elucidate the role of PRPF8/Brr2 regulatory mechanisms in splicing and the molecular basis of retinal disease, informing therapeutic approaches.
(© 2024. The Author(s).)
Databáze: MEDLINE
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