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PLoS One
2014 Jan 01;94:e95768. doi: 10.1371/journal.pone.0095768.
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A truncated form of rod photoreceptor PDE6 β-subunit causes autosomal dominant congenital stationary night blindness by interfering with the inhibitory activity of the γ-subunit.
Manes G
,
Cheguru P
,
Majumder A
,
Bocquet B
,
Sénéchal A
,
Artemyev NO
,
Hamel CP
,
Brabet P
.
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Autosomal dominant congenital stationary night blindness (adCSNB) is caused by mutations in three genes of the rod phototransduction cascade, rhodopsin (RHO), transducin α-subunit (GNAT1), and cGMP phosphodiesterase type 6 β-subunit (PDE6B). In most cases, the constitutive activation of the phototransduction cascade is a prerequisite to cause adCSNB. The unique adCSNB-associated PDE6B mutation found in the Rambusch pedigree, the substitution p.His258Asn, leads to rod photoreceptors desensitization. Here, we report a three-generation French family with adCSNB harboring a novel PDE6B mutation, the duplication, c.928-9_940dup resulting in a tyrosine to cysteine substitution at codon 314, a frameshift, and a premature termination (p.Tyr314Cysfs*50). To understand the mechanism of the PDE6β1-314fs*50 mutant, we examined the properties of its PDE6-specific portion, PDE6β1-313. We found that PDE6β1-313 maintains the ability to bind noncatalytic cGMP and the inhibitory γ-subunit (Pγ), and interferes with the inhibition of normal PDE6αβ catalytic subunits by Pγ. Moreover, both truncated forms of the PDE6β protein, PDE6β1-313 and PDE6β1-314fs*50 expressed in rods of transgenic X. laevis are targeted to the phototransduction compartment. We hypothesize that in affected family members the p.Tyr314Cysfs*50 change results in the production of the truncated protein, which binds Pγ and causes constitutive activation of the phototransduction thus leading to the absence of rod adaptation.
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Figure 2. Clinical investigations of the index patient (III:1) of a French family with adCSNB.A. Fundus photographs of index patient III:1, right eye and left eye. B. Dark-adaptation curve of index patient III:1 (solid line) compared to normal curve (hatched line). D. Full-field ERGs from a normal control and index patient III:1.
Figure 3. Interaction of PDE6β1-313 with the polycationic region of Pγ, Pγ24-45.A. Emission spectra of CysBC-Pγ24-45 (50 nM) alone (1) and in the presence of PDE6β1-313 (180 nM) (2) were recorded with with excitation at 445 nm. Inset: Coomassie Blue-stained gel with the sample of PDE6β1-313. B. Fluorescence of CysBC-Pγ24-45 (50 nM) was measured on addition of increasing concentrations of PDE6β1-313 (excitation at 445 nm, emission at 490 nm). The data were fit to the hyperbola equation using GraphPad Prizm software. C. Noncatalytic cGMP binding to PDE6β1-313. Binding of cGMP was carried out using [H3]cGMP and varying concentrations of unlabeled cGMP. The bound cGMP was determined by the filter-binding assay, and is expressed as a percentage of maximal cGMP binding (0.5 mol cGMP/mol PDE6β1-313). Results from one of three similar experiments are shown. The data were fit to equation for binding with ligand depletion using GraphPad Prizm software. The calculated Kd = 0.82±0.15 µM.
Figure 4. Effect of PDE6β1-313 on the inhibition of trypsin-activated PDE6 (tPDE6) by Pγ.The activity of tPDE6 was determined upon addition of increasing concentrations of Pγ in the absence (squares) or in the presence (circles) of 2 µM PDE6β1-313. The tPDE6 activity is plotted as a function of Pγ concentration. Results from one of three similar experiments are shown. The data were fit to sigmoidal dose-response equation using GraphPad Prizm Software. The calculated IC50 values (squares, 0.53±0.07 nM; circles, 1.6±0.2 nM) are significantly different (p<0.01).
Figure 5. Expression of PDE6β1-314fs*50 in transgenic X. laevis rods photoreceptors.A. left - EGFP fluorescence in living photoreceptor cells expressing PDE6β1-314fs*50, center - DIC image, right - EGFP-fluorescence/DIC overlay. Bar - 10 µm. B. Co-localization of PDE6β1-314fs*50 (green) with the rod outer segment marker Wheat Germ Agglutinin (red) in cryosections of transgenic retina. Cryosections were counterstained with TO-PRO3 nuclear stain (blue). Bar - 10 µm. C. Isotonic (I), hypotonic (H) extracts of transgenic PDE6β1-314fs*50 retinas were analyzed by immunoblotting with anti-GFP B-2 monoclonal antibody.
Figure 6. Expression of EGFP-PDE6β1-313 in transgenic X. laevis rods photoreceptors.A. left - EGFP fluorescence in living photoreceptor cells expressing EGFP-PDE6β1-313, center - DIC image, right - EGFP-fluorescence/DIC overlay. Bar - 10 µm. B. Co-localization of EGFP-PDE6β1-313 (green) with the rod outer segment marker Wheat Germ Agglutinin (red) in cryosections of transgenic retina. Cryosections were counterstained with TO-PRO3 nuclear stain (blue). Bar - 10 µm. C. Isotonic (I), hypotonic (H) extracts of transgenic EGFP-PDE6β1-313 retinas were analyzed by immunoblotting with anti-GFP B-2 monoclonal antibody.
Figure 1. Pedigree with adCSNB and PDE6B mutation.A. The three-generation French family MTP1481 with adCSNB. Filled symbols indicate affected family members; squares: males; circles: females; arrow: index patient; slashed symbols: deceased persons; question marks: family members with no information about the phenotype available. M indicates the presence of the mutation and + indicate the wild-type allele. B. Electropherograms show the normal control sequence and affected sequence (index individual III:1) surrounding the p.Tyr314Cysfs*50 mutation. The dotted line represents the 22 duplicated nucleotides. C. Schematic representation of the PDE6β protein showing the domain structure and positions of arRP truncating mutations (below) and adCSNB mutations (above) including the mutation described in this study (Y314Cfs*50).
Aquirre,
Rod-cone dysplasia in Irish setters: a defect in cyclic GMP metabolism in visual cells.
1978, Pubmed
Aquirre,
Rod-cone dysplasia in Irish setters: a defect in cyclic GMP metabolism in visual cells.
1978,
Pubmed
Arshavsky,
Photoreceptor signaling: supporting vision across a wide range of light intensities.
2012,
Pubmed
Artemyev,
Two-site high-affinity interaction between inhibitory and catalytic subunits of rod cyclic GMP phosphodiesterase.
1992,
Pubmed
Baehr,
Isolation and characterization of cGMP phosphodiesterase from bovine rod outer segments.
1979,
Pubmed
Chabre,
Molecular mechanism of visual transduction.
1989,
Pubmed
Cheguru,
The GAFa domain of phosphodiesterase-6 contains a rod outer segment localization signal.
2014,
Pubmed
,
Xenbase
Danciger,
Mutations in the PDE6B gene in autosomal recessive retinitis pigmentosa.
1995,
Pubmed
Danciger,
A homozygous PDE6B mutation in a family with autosomal recessive retinitis pigmentosa.
1996,
Pubmed
Deterre,
cGMP phosphodiesterase of retinal rods is regulated by two inhibitory subunits.
1988,
Pubmed
Dryja,
Heterozygous missense mutation in the rhodopsin gene as a cause of congenital stationary night blindness.
1993,
Pubmed
Dryja,
Missense mutation in the gene encoding the alpha subunit of rod transducin in the Nougaret form of congenital stationary night blindness.
1996,
Pubmed
Dryja,
Molecular genetics of Oguchi disease, fundus albipunctatus, and other forms of stationary night blindness: LVII Edward Jackson Memorial Lecture.
2000,
Pubmed
Dvir,
Autosomal-recessive early-onset retinitis pigmentosa caused by a mutation in PDE6G, the gene encoding the gamma subunit of rod cGMP phosphodiesterase.
2010,
Pubmed
Farber,
Cyclic guanosine monophosphate: elevation in degenerating photoreceptor cells of the C3H mouse retina.
1974,
Pubmed
Farber,
Enzymic basis for cyclic GMP accumulation in degenerative photoreceptor cells of mouse retina.
1976,
Pubmed
Frischmeyer,
Nonsense-mediated mRNA decay in health and disease.
1999,
Pubmed
Gal,
Heterozygous missense mutation in the rod cGMP phosphodiesterase beta-subunit gene in autosomal dominant stationary night blindness.
1994,
Pubmed
Granovsky,
Probing domain functions of chimeric PDE6alpha'/PDE5 cGMP-phosphodiesterase.
1998,
Pubmed
Guo,
The inhibitory gamma subunit of the rod cGMP phosphodiesterase binds the catalytic subunits in an extended linear structure.
2006,
Pubmed
Holbrook,
Nonsense-mediated decay approaches the clinic.
2004,
Pubmed
Huang,
Autosomal recessive retinitis pigmentosa caused by mutations in the alpha subunit of rod cGMP phosphodiesterase.
1995,
Pubmed
Hurley,
Purification and characterization of the gamma regulatory subunit of the cyclic GMP phosphodiesterase from retinal rod outer segments.
1982,
Pubmed
Karan,
A model for transport of membrane-associated phototransduction polypeptides in rod and cone photoreceptor inner segments.
2008,
Pubmed
Kroll,
Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation.
1996,
Pubmed
,
Xenbase
Lolley,
Cyclic GMP accumulation causes degeneration of photoreceptor cells: simulation of an inherited disease.
1977,
Pubmed
,
Xenbase
Majumder,
Interaction of aryl hydrocarbon receptor-interacting protein-like 1 with the farnesyl moiety.
2013,
Pubmed
Martinez,
The structure of the GAF A domain from phosphodiesterase 6C reveals determinants of cGMP binding, a conserved binding surface, and a large cGMP-dependent conformational change.
2008,
Pubmed
McLaughlin,
Recessive mutations in the gene encoding the beta-subunit of rod phosphodiesterase in patients with retinitis pigmentosa.
1993,
Pubmed
McLaughlin,
Mutation spectrum of the gene encoding the beta subunit of rod phosphodiesterase among patients with autosomal recessive retinitis pigmentosa.
1995,
Pubmed
Miller,
A simple salting out procedure for extracting DNA from human nucleated cells.
1988,
Pubmed
Moussaif,
Phototransduction in a transgenic mouse model of Nougaret night blindness.
2006,
Pubmed
Muradov,
Loss of the effector function in a transducin-alpha mutant associated with Nougaret night blindness.
2000,
Pubmed
Muradov,
Direct interaction of the inhibitory gamma-subunit of Rod cGMP phosphodiesterase (PDE6) with the PDE6 GAFa domains.
2002,
Pubmed
Muradov,
Mutation in rod PDE6 linked to congenital stationary night blindness impairs the enzyme inhibition by its gamma-subunit.
2003,
Pubmed
Muradov,
Structural determinants of the PDE6 GAF A domain for binding the inhibitory gamma-subunit and noncatalytic cGMP.
2004,
Pubmed
Muradov,
PDE6 in lamprey Petromyzon marinus: implications for the evolution of the visual effector in vertebrates.
2007,
Pubmed
Muradov,
Characterization of human cone phosphodiesterase-6 ectopically expressed in Xenopus laevis rods.
2009,
Pubmed
,
Xenbase
Naeem,
GNAT1 associated with autosomal recessive congenital stationary night blindness.
2012,
Pubmed
Natochin,
An interface of interaction between photoreceptor cGMP phosphodiesterase catalytic subunits and inhibitory gamma subunits.
1996,
Pubmed
Papermaster,
Rhodopsin content in the outer segment membranes of bovine and frog retinal rods.
1974,
Pubmed
Rao,
Rhodopsin mutation G90D and a molecular mechanism for congenital night blindness.
1994,
Pubmed
Singhal,
Insights into congenital stationary night blindness based on the structure of G90D rhodopsin.
2013,
Pubmed
Suber,
Irish setter dogs affected with rod/cone dysplasia contain a nonsense mutation in the rod cGMP phosphodiesterase beta-subunit gene.
1993,
Pubmed
Szabo,
p.Gln200Glu, a putative constitutively active mutant of rod alpha-transducin (GNAT1) in autosomal dominant congenital stationary night blindness.
2007,
Pubmed
Ulshafer,
Sensitivity of photoreceptors to elevated levels of cGMP in the human retina.
1980,
Pubmed
Yarfitz,
Transduction mechanisms of vertebrate and invertebrate photoreceptors.
1994,
Pubmed
Zeitz,
Identification and functional characterization of a novel rhodopsin mutation associated with autosomal dominant CSNB.
2008,
Pubmed
al-Jandal,
A novel mutation within the rhodopsin gene (Thr-94-Ile) causing autosomal dominant congenital stationary night blindness.
1999,
Pubmed
