Three novel rhodopsin mutations (C110F, L131P, A164V) in patients with autosomal dominant retinitis pigmentosa

Fuchs, Sigrid; Kranich, Holger; Denton, M. J.; Zrenner, Eberhart; Bhattacharya, SS; Humphries, Peter; Gál, A.

doi:10.1093/hmg/3.7.1203

Cited by 26 publications

(16 citation statements)

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“…34,35 Disease-causing mutations of the highly conserved disulfide bond connecting ECL1 and ECL2 have been identified in many GPCRs, including AVPR2, 36 RHO 37,38 and MC2R. 39 About 7% of all missense mutations in GPCR are substitutions with cysteine residues found in the putative N-terminal portion and ECL1-3.…”

Section: Discussionmentioning

confidence: 99%

Identification of two new mutations in the GPR98 and the PDE6B genes segregating in a Tunisian family

Hmani-Aifa¹,

Benzina

Zulfiqar

et al. 2008

Eur J Hum Genet

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Autosomal recessive retinitis pigmentosa (ARRP) is a genetically heterogeneous disorder. ARRP could be associated with extraocular manifestations that define specific syndromes such as Usher syndrome (USH) characterized by retinal degeneration and congenital hearing loss (HL). The USH type II (USH2) associates RP and mild-to-moderate HL with preserved vestibular function. At least three genes USH2A, the very large G-protein-coupled receptor, GPR98, and DFNB31 are responsible for USH2 syndrome. Here, we report on the segregation of non-syndromic ARRP and USH2 syndrome in a consanguineous Tunisian family, which was previously used to define USH2B locus. With regard to the co-occurrence of these two different pathologies, clinical and genetic reanalysis of the extended family showed (i) phenotypic heterogeneity within USH2 patients and (ii) excluded linkage to USH2B locus. Indeed, linkage analysis disclosed the cosegregation of the USH2 phenotype with the USH2C locus markers, D5S428 and D5S618, whereas the ARRP perfectly segregates with PDE6B flanking markers D4S3360 and D4S2930. Molecular analysis revealed two new missense mutations, p.Y6044C and p.W807R, occurring in GPR98 and PDE6B genes, respectively. In conclusion, our results show that the USH2B locus at chromosome 3p23 -24.2 does not exist, and we therefore withdraw this locus designation. The combination of molecular findings for GPR98 and PDE6B genes enable us to explain the phenotypic heterogeneity and particularly the severe ocular affection first observed in one USH2 patient. This report presents an illustration of how consanguinity could increase familial clustering of multiple hereditary diseases within the same family.

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Section: Discussionmentioning

confidence: 99%

Identification of two new mutations in the GPR98 and the PDE6B genes segregating in a Tunisian family

Hmani-Aifa¹,

Benzina

Zulfiqar

et al. 2008

Eur J Hum Genet

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“…This result paralleled the earlier findings on the small-protein bovine pancreatic trypsin inhibitor, and, by analogy, suggested a globular structure for the intradiscal domain in rhodopsin (3). Recently, point mutations at the intradiscal cysteines (C110F, C110Y, and C187Y) in rhodopsin associated with RP have been discovered (14)(15)(16)(17) and, in addition, mutation of a conserved cysteine equivalent to Cys-187 in cone opsins (C203R) has been shown to cause color vision deficiencies (18,19).…”

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confidence: 99%

Structure and function in rhodopsin: Further elucidation of the role of the intradiscal cysteines, Cys-110, -185, and -187, in rhodopsin folding and function

Hwa

Reeves

Klein‐Seetharaman

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

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The disulfide bond between Cys-110 and Cys-187 in the intradiscal domain is required for correct folding in vivo and function of mammalian rhodopsin. Misfolding in rhodopsin, characterized by the loss of ability to bind 11-cis-retinal, has been shown to be caused by an intradiscal disulfide bond different from the above native disulfide bond. Further, naturally occurring single mutations of the intradiscal cysteines (C110F, C110Y, and C187Y) are associated with retinitis pigmentosa (RP). To elucidate further the role of every one of the three intradiscal cysteines, mutants containing single-cysteine replacements by alanine residues and the above three RP mutants have been studied. We find that C110A, C110F, and C110Y all form a disulfide bond between C185 and C187 and cause loss of retinal binding. C185A allows the formation of a C110-C187 disulfide bond, with wild-type-like rhodopsin phenotype. C187A forms a disulfide bond between C110 and C185 and binds retinal, and the pigment formed has markedly altered bleaching behavior. However, the opsin from the RP mutant C187Y forms no rhodopsin chromophore.Early work indicated the important role of the intradiscal domain in folding in vivo and function of rhodopsin (1). The presence of a disulfide bond between Cys-110 and Cys-187 was established (2) and it was shown later to be required for optimal signal transduction (3). A disulfide bond between two cysteines in the extracellular domain at positions equivalent to the above two cysteines in rhodopsin is known now to be conserved in most of the known G-protein-coupled receptors (4, 5). The presence of a disulfide bond indicated a tertiary structure in the intradiscal domain of rhodopsin. Strong support for this conclusion was forthcoming from extensive designed mutagenic studies as well as from studies of a large number of naturally occurring point mutations in rhodopsin that are associated with retinitis pigmentosa (RP) (6-11). Mutations in the intradiscal domain were shown to cause total or partial misfolding in vivo in rhodopsin, misfolding being operationally defined as the loss of ability to bind 11-cis retinal (6). Further, evidence was presented that misfolding results from the formation of a disulfide bond in the intradiscal domain different from the normal Cys-110-Cys-187 disulfide bond (12). Recently, Oprian and colleagues demonstrated the formation of a disulfide bond under certain conditions between Cys-185 and Cys-187 in a rhodopsin mutant reconstituted from two appropriate rhodopsin fragments (13).Further studies showed that although replacements of Cys-110 and Cys-187 by serine residues prevented folding of rhodopsin to a functional structure, corresponding replacements by alanine residues allowed the formation of the correctly folded dark-state structure. This result paralleled the earlier findings on the small-protein bovine pancreatic trypsin inhibitor, and, by analogy, suggested a globular structure for the intradiscal domain in rhodopsin (3). Recently, point mutations at the intradiscal cys...

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“…We show by kinetic studies of retinal binding that the mutants show large variation in rates of retinal binding. We have chosen as examples the tryptophan 265 mutants, W265F,Y,A (7), as well as the retinitis pigmentosa (RP) mutant A164V (9,10) in the transmembrane domain (Fig. 1).…”

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confidence: 99%

Structure and function in rhodopsin: Kinetic studies of retinal binding to purified opsin mutants in defined phospholipid–detergent mixtures serve as probes of the retinal binding pocket

Reeves

Hwa²,

Khorana³

1999

Proc. Natl. Acad. Sci. U.S.A.

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In the current standard procedure for preparation of mammalian rhodopsin mutants, transfected COS-1 cells expressing the mutant opsin genes are treated with 5 M 11-cis-retinal before detergent solubilization for purification. We found that binding of 11-cis-retinal to opsin mutants with single amino acid changes at Trp-265 (W265F,Y,A) and a retinitis pigmentosa mutant (A164V) was far from complete and required much higher concentrations of 11-cis-retinal. By isolation of the expressed opsins in a stable form, kinetic studies of retinal binding to the opsins in vitro have been carried out by using defined phospholipid-detergent mixtures. The results show wide variation in the rates of 11-cisretinal binding. Thus, the in vitro reconstitution procedure serves as a probe of the retinal-binding pocket in the opsins. Further, a method is described for purification and characterization of the rhodopsin mutants after retinal binding to the opsins in vitro.

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Three novel rhodopsin mutations (C110F, L131P, A164V) in patients with autosomal dominant retinitis pigmentosa

Cited by 26 publications

References 0 publications

Identification of two new mutations in the GPR98 and the PDE6B genes segregating in a Tunisian family

Identification of two new mutations in the GPR98 and the PDE6B genes segregating in a Tunisian family

Structure and function in rhodopsin: Further elucidation of the role of the intradiscal cysteines, Cys-110, -185, and -187, in rhodopsin folding and function

Structure and function in rhodopsin: Kinetic studies of retinal binding to purified opsin mutants in defined phospholipid–detergent mixtures serve as probes of the retinal binding pocket

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