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A novel missense mutation of RPGR identified from retinitis pigmentosa affects splicing of the ORF15 region and causes loss of transcript heterogeneity
- Source :
- Biochemical and Biophysical Research Communications. 531:172-179
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene, are the major cause of X-linked retinitis pigmentosa (RP), in which exon open reading frame 15 (ORF15) of RPGR has been implicated to play a substantial role. We identified a novel hemizygous missense mutation E585K of RPGR from whole-exome sequencing of RP. RNA-Seq analysis and functional study were conducted to investigate the underlying pathogenic mechanism of the mutation. Our results showed that the mutation actually affected RPGR ORF15 splicing. RNA-Seq analysis of the human retina followed by validation in cells revealed a complex splicing pattern near the 3′ boundary of RPGR exon 14 in the ORF15 region, resulting from a variety of alternative splicing events (ASEs). The wildtype RPGR mini-gene expressed in human 293T cells confirmed these ASEs in vitro. In contrast, without new RNA species detected, the mutant mini-gene disrupted the splicing pattern of the ORF15 region, and caused loss of RPGR transcript heterogeneity. The RNA species derived from the mutant mini-gene were predominated by a minor out-of-frame transcript that was also observed in wildtype RPGR, resulting from an upstream alternative 5′ splice site in exon 14. Our findings therefore provide insights into the influence of RPGR exonic mutations on alternative splicing of the ORF15 region, and the underlying molecular mechanism of RP.
- Subjects :
- Male
0301 basic medicine
RNA Splicing
Mutation, Missense
Biophysics
Biology
medicine.disease_cause
Biochemistry
Cell Line
Open Reading Frames
03 medical and health sciences
Exon
0302 clinical medicine
Retinitis pigmentosa
medicine
Humans
Missense mutation
Amino Acid Sequence
RNA, Messenger
Eye Proteins
Molecular Biology
Gene
Hemizygote
Genetics
Mutation
Base Sequence
Alternative splicing
Cell Biology
Retinitis pigmentosa GTPase regulator
medicine.disease
eye diseases
030104 developmental biology
030220 oncology & carcinogenesis
RNA splicing
Retinitis Pigmentosa
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 531
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....a75c063039bf48d99a73eacbaab06453