Disruption of RPGR protein interaction network is the common feature of RPGR missense variations that cause XLRP

Qihong Zhang; Joseph C Giacalone; Charles Searby; Edwin M Stone; Budd A Tucker; Val C Sheffield

doi:10.1073/pnas.1817639116

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Disruption of RPGR protein interaction network is the common feature of RPGR missense variations that cause XLRP

Journal article

Open access

Peer reviewed

Disruption of RPGR protein interaction network is the common feature of RPGR missense variations that cause XLRP

Qihong Zhang, Joseph C Giacalone, Charles Searby, Edwin M Stone, Budd A Tucker and Val C Sheffield

Proceedings of the National Academy of Sciences - PNAS, Vol.116(4), pp.1353-1360

01/22/2019

DOI: 10.1073/pnas.1817639116

PMCID: PMC6347721

PMID: 30622176

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Abstract

Retinitis pigmentosa (RP) is an inherited retinal degenerative disease with severe vision impairment leading to blindness. About 10-15% of RP cases are caused by mutations in the gene, with RPGR mutations accounting for 70% of X-linked RP cases. The mechanism by which RPGR mutations cause photoreceptor cell dysfunction is not well understood. In this study, we show that the two isoforms of RPGR (RPGR and RPGR ) interact with endogenous PDE6D, INPP5E, and RPGRIP1L. The RPGR isoform contains two PDE6D binding sites with the C-terminal prenylation site being the predominant PDE6D binding site. The C terminus of RPGR that contains the prenylation site regulates its interaction with PDE6D, INPP5E, and RPGRIP1L. Only the RPGR isoform localizes to cilia in cultured RPE1 cells. Missense variations found in RPGR patients disrupt the interaction between RPGR isoforms and their endogenous interactors INPP5E, PDE6D, and RPGRIP1L. We evaluated a RPGR missense variation (M58K) found in a family with X-linked retinitis pigmentosa (XLRP) and show that this missense variation disrupts the interaction of RPGR isoforms with their endogenous interactors. The M58K variation also disrupts the ciliary localization of the RPGR isoform. Using this assay, we also show that some of the RPGR missense variants reported in the literature might not actually be disease causing. Our data establishes an in vitro assay that can be used to validate the potential pathogenicity of RPGR missense variants.

Cell Line

Humans

Retinitis Pigmentosa - genetics

Binding Sites - genetics

Mutation, Missense - genetics

Cilia - genetics

Animals

Protein Interaction Domains and Motifs - genetics

HEK293 Cells

Mice

Eye Proteins - genetics

Nuclear Proteins - genetics

Protein Isoforms - genetics

Details

Title: Subtitle: Disruption of RPGR protein interaction network is the common feature of RPGR missense variations that cause XLRP
Creators: Qihong Zhang - Department of Pediatrics, University of Iowa, Iowa City, IA 52242; qihong-zhang@uiowa.edu val-sheffield@uiowa.edu
Joseph C Giacalone - Department of Ophthalmology and Visual Sciences, University of Iowa Carver College of Medicine, Iowa City, IA 52242
Charles Searby - Department of Pediatrics, University of Iowa, Iowa City, IA 52242
Edwin M Stone - Institute for Vision Research, University of Iowa, Iowa City, IA 52242
Budd A Tucker - Institute for Vision Research, University of Iowa, Iowa City, IA 52242
Val C Sheffield - Institute for Vision Research, University of Iowa, Iowa City, IA 52242
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.116(4), pp.1353-1360
DOI: 10.1073/pnas.1817639116
PMID: 30622176
PMCID: PMC6347721
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; United States
Grant note: R01 EY011298 / NEI NIH HHS P30 EY025580 / NEI NIH HHS R01 EY026008 / NEI NIH HHS T32 GM007337 / NIGMS NIH HHS R01 EY017168 / NEI NIH HHS
Language: English
Date published: 01/22/2019
Academic Unit: Stead Family Department of Pediatrics; The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; Medical Genetics and Genomics; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Ophthalmology and Visual Sciences
Record Identifier: 9984070224302771

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