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

Autor: Atkinson, Robert, Georgiou, Maria, Chunbo Yang, Szymanska, Katarzyna, Lahat, Albert, Vasconcelos, Elton J. R., Yanlong Ji, Molina, Marina Moya, Collin, Joseph, Queen, Rachel, Dorgau, Birthe, Watson, Avril, Kurzawa-Akanbi, Marzena, Laws, Ross, Saxena, Abhijit, Beh, Chia Shyan, Siachisumo, Chileleko, Goertler, Franziska, Karwatka, Magdalena, Davey, Tracey
Zdroj: Nature Communications; 4/11/2024, Vol. 15 Issue 1, p1-17, 17p, 8 Graphs
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 retinalspecific 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. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
Externí odkaz: