Browse by author
Lookup NU author(s): Rob Atkinson, Dr Maria Georgiou, Dr Chunbo Yang, Marina Moya Molina, Dr Joseph Collin, Dr Rachel Queen, Avril Watson, Dr Marzena Kurzawa-Akanbi, Ross LawsORCiD, Abhijit Saxena, Chia Shyan Beh Beh, Chileleko Siachisumo, Tracey DaveyORCiD, Professor David SteelORCiD, Professor David Elliott, Professor Lyle Armstrong, Professor Majlinda LakoORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© The Author(s) 2024.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.
Author(s): Atkinson R, Georgiou M, Yang C, Szymanska K, Lahat A, Vasconcelos EJR, Ji Y, Moya Molina M, Collin J, Queen R, Dorgau B, Watson A, Kurzawa-Akanbi M, Laws R, Saxena A, Shyan Beh C, Siachisumo C, Goertler F, Karwatka M, Davey T, Inglehearn CF, McKibbin M, Luhrmann R, Steel DH, Elliott DJ, Armstrong L, Urlaub H, Ali RR, Grellscheid S-N, Johnson CA, Mozaffari-Jovin S, Lako M
Publication type: Article
Publication status: Published
Journal: Nature Communications
Year: 2024
Volume: 15
Issue: 1
Online publication date: 11/04/2024
Acceptance date: 19/03/2024
Date deposited: 18/04/2024
ISSN (electronic): 2041-1723
Publisher: Nature Research
URL: https://doi.org/10.1038/s41467-024-47253-0
DOI: 10.1038/s41467-024-47253-0
Data Access Statement: The trimmed FASTQ data for bulk RNA-Seq of all samples included in this study were uploaded to SRA under the accession number BioProject ID PRJNA989762 and GEO under the accession number GSE236702. The mass spectrometry proteomics data have been deposited to the ProteomeXchange under the accession number PXD043645. The single cell RNA-Seq data have been deposited to the GEO under the accession number GSE235866. The iCLIP-Seq data have been uploaded to Annotare under the accession number E-MTAB13171. A source data file has been provided with the manuscript. Source data are provided in this paper. No new codes were used in this study.
Altmetrics provided by Altmetric
