Retinitis pigmentosa-linked mutation in DHX38 modulates its splicing activity

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 2nd step of splicing. We showed that D...

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Bibliographic Details
Published in:PloS one 2022-04, Vol.17 (4), p.e0265742-e0265742
Main Authors: Obuca, Mina, Kubovciak, Jan, Stanek, David, Cvacková, Zuzana, Kolár, Michal
Format: Article
Language:English
Subjects:
Alternative splicing
Amino acid substitution
Amino acids
Analysis
Biology and life sciences
DEAD-box RNA Helicases - genetics
Diagnosis
DNA helicase
Efficiency
Gene expression
Genes
Genetic aspects
Genetic disorders
Hereditary diseases
Humans
Medicine and Health Sciences
Messenger RNA
Mutation
Perturbation
Plasmids
Proteins
Research and Analysis Methods
Retina
Retinitis
Retinitis pigmentosa
Retinitis Pigmentosa - genetics
RNA helicase
RNA Splice Sites
RNA Splicing
RNA Splicing Factors - genetics
Spliceosomes - metabolism
Splicing factors
Substitutes
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Summary: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 2nd 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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0265742