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

Atkinson, Robert; Georgiou, Maria; Yang, Chunbo; Szymanska, Katarzyna; Lahat, Albert; Vasconcelos, Elton J R; Ji, Yanlong; Moya Molina, Marina; Collin, Joseph; Queen, Rachel; Dorgau, Birthe; Watson, Avril; Kurzawa-Akanbi, Marzena; Laws, Ross; Saxena, Abhijit; Shyan Beh, Chia; Siachisumo, Chileleko; Goertler, Franziska; Karwatka, Magdalena; Davey, Tracey; Inglehearn, Chris F; McKibbin, Martin; Lührmann, Reinhard; Steel, David H; Elliott, David J; Armstrong, Lyle; Urlaub, Henning; Ali, Robin R; Grellscheid, Sushma-Nagaraja; Johnson, Colin A; Mozaffari-Jovin, Sina; Lako, Majlinda

doi:10.1038/s41467-024-47253-0

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

Atkinson, Robert; Georgiou, Maria; Yang, Chunbo; Szymanska, Katarzyna; Lahat, Albert; Vasconcelos, Elton J R; Ji, Yanlong; Moya Molina, Marina; Collin, Joseph; Queen, Rachel; Dorgau, Birthe; Watson, Avril; Kurzawa-Akanbi, Marzena; Laws, Ross; Saxena, Abhijit; Shyan Beh, Chia; Siachisumo, Chileleko; Goertler, Franziska; Karwatka, Magdalena; Davey, Tracey; Inglehearn, Chris F; McKibbin, Martin; Lührmann, Reinhard; Steel, David H; Elliott, David J; Armstrong, Lyle; Urlaub, Henning; Ali, Robin R; Grellscheid, Sushma-Nagaraja; Johnson, Colin A; Mozaffari-Jovin, Sina; Lako, Majlinda

Authors

Robert Atkinson

Maria Georgiou

Chunbo Yang

Katarzyna Szymanska

Dr Albert Lahat albert.lahat@durham.ac.uk
Post Doctoral Research Associate

Elton J R Vasconcelos

Yanlong Ji

Marina Moya Molina

Joseph Collin

Rachel Queen

Birthe Dorgau

Avril Watson

Marzena Kurzawa-Akanbi

Ross Laws

Abhijit Saxena

Chia Shyan Beh

Chileleko Siachisumo

Franziska Goertler

Magdalena Karwatka

Tracey Davey

Chris F Inglehearn

Martin McKibbin

Reinhard Lührmann

David H Steel

David J Elliott

Lyle Armstrong

Henning Urlaub

Robin R Ali

Professor Sushma Grellscheid s.n.grellscheid@durham.ac.uk
Professor

Colin A Johnson

Sina Mozaffari-Jovin

Majlinda Lako

Abstract

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.

Citation

Atkinson, R., Georgiou, M., Yang, C., Szymanska, K., Lahat, A., Vasconcelos, E. J. R., …Lako, M. (2024). PRPF8-mediated dysregulation of hBrr2 helicase disrupts human spliceosome kinetics and 5´-splice-site selection causing tissue-specific defects. Nature Communications, 15(1), Article 3138. https://doi.org/10.1038/s41467-024-47253-0

Journal Article Type	Article
Acceptance Date	Mar 19, 2024
Online Publication Date	Apr 11, 2024
Publication Date	2024-04
Deposit Date	May 15, 2024
Publicly Available Date	May 15, 2024
Journal	Nature communications
Electronic ISSN	2041-1723
Publisher	Nature Research
Peer Reviewed	Peer Reviewed
Volume	15
Issue	1
Article Number	3138
DOI	https://doi.org/10.1038/s41467-024-47253-0
Keywords	Spliceosomes, Humans, Retinitis Pigmentosa, DNA Helicases, RNA-Binding Proteins, RNA, Messenger, RNA Splice Sites, RNA, Small Nuclear, Proteomics, RNA Splicing, Alternative Splicing, Mutation
Public URL	https://durham-repository.worktribe.com/output/2407261
PMID	38605034

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