Retinitis pigmentosa-linked mutation in DHX38 modulates its splicing activity

0558860 - ÚMG 2023 RIV US eng J - Journal Article
Obuca, Mina - Cvačková, Zuzana - Kubovčiak, Jan - Kolář, Michal - Staněk, David
Retinitis pigmentosa-linked mutation in DHX38 modulates its splicing activity.
PLoS ONE. Roč. 17, č. 4 (2022), č. článku e0265742. ISSN 1932-6203. E-ISSN 1932-6203
R&D Projects: GA ČR(CZ) GC18-01911J; GA MŠMT(CZ) EF16_019/0000785
Institutional support: RVO:68378050
Keywords : pre-messenger-rna * atpase prp16 * helix i * spliceosome * mechanism * fidelity * impairs * gene
OECD category: Cell biology
Impact factor: 3.7, year: 2022
Method of publishing: Open access
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0265742

Retinitis pigmentosa (RP) is a hereditary disease affecting tens of thousands of people world-wide. Here we analyzed the effect of an amino acid substitution in the RNA helicase DHX38 (Prp16) causing RP. DHX38 has been proposed as the helicase important for the 2(nd) step of splicing. We showed that DHX38 associates with key splicing factors involved in both splicing steps but did not find any evidence that the RP mutations changes DHX38 interaction profile with the spliceosome. We further downregulated DHX38 and monitored changes in splicing. We observed only minor perturbations of general splicing but detected modulation of ~70 alternative splicing events. Next, we probed DHX38 function in splicing of retina specific genes and found that FSCN2 splicing is dependent on DHX38. In addition, RHO splicing was inhibited specifically by expression of DHX38 RP variant. Finally, we showed that overexpression of DHX38 promotes usage of canonical as well as cryptic 5' splice sites in HBB splicing reporter. Together, our data show that DHX38 is a splicing factor that promotes splicing of cryptic splice sites and regulate alternative splicing. We further provide evidence that the RP-linked substitution G332D modulates DHX38 splicing activity.
Permanent Link: https://hdl.handle.net/11104/0332899