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Mutations in the rhodopsin gene cause approximately one-tenth of retinitis pigmentosa cases worldwide, and most result in endoplasmic reticulum retention and apoptosis. Other rhodopsin mutations cause receptor mislocalization, diminished/constitutive activity, or faulty protein-protein interactions. The purpose of this study was to test for mechanisms by which the autosomal dominant rhodopsin mutation Ter349Glu causes an early, rapid retinal degeneration in patients. The mutation adds an additional 51 amino acids to the C terminus of the protein. Folding and ligand interaction of Ter349Glu rhodopsin were tested by ultraviolet-visible (UV-visible) spectrophotometry. The ability of the mutant to initiate phototransduction was tested using a radioactive filter binding assay. Photoreceptor localization was assessed both in vitro and in vivo utilizing fluorescent immunochemistry on transfected cells, transgenic Xenopus laevis, and knock-in mice. Photoreceptor ultrastructure was observed by transmission electron microscopy. Spectrally, Ter349Glu rhodopsin behaves similarly to wild-type rhodopsin, absorbing maximally at 500 nm. The mutant protein also displays in vitro G protein activation similar to that of WT. In cultured cells, mislocalization was observed at high expression levels whereas ciliary localization occurred at low expression levels. Similarly, transgenic X. laevis expressing Ter349Glu rhodopsin exhibited partial mislocalization. Analysis of the Ter349Glu rhodopsin knock-in mouse showed a rapid, early onset degeneration in homozygotes with a loss of proper rod outer segment development and improper disc formation. Together, the data show that both mislocalization and rod outer segment morphogenesis are likely associated with the human phenotype.
FIGURE 3. Localization of Ter349Glu rhodopsin in transgenic X. laevis.
Using a modified Amaya and Kroll method of transgenesis,WTand Ter349Glu
rhodopsin-expressing tadpoles were generated and euthanized at 2 weeks of
age, processed and cryosectioned, and labeled using immunohistochemistry.
WT rhodopsin was labeled using B6-30N primary antibody. Ter349Glu
rhodopsin was labeled using mammalian rhodopsin-specific primary antibody
A5-3-12. Both were labeled with anti-mouse IgG secondary antibody
conjugated to Alexa Fluor 488 (green). Nuclei were labeled with DAPI (blue).
OPL, outer plexiform layer. Arrows, normally developed outer segments;
arrowheads (>), inner segment mislocalization. Asterisks, synaptic mislocalization.
Scale bar, 20 m.
Abd-El-Barr,
Impaired photoreceptor protein transport and synaptic transmission in a mouse model of Bardet-Biedl syndrome.
2007, Pubmed
Abd-El-Barr,
Impaired photoreceptor protein transport and synaptic transmission in a mouse model of Bardet-Biedl syndrome.
2007,
Pubmed
al-Maghtheh,
Rhodopsin mutations in autosomal dominant retinitis pigmentosa.
1993,
Pubmed
Amaya,
A method for generating transgenic frog embryos.
1999,
Pubmed
,
Xenbase
Apfelstedt-Sylla,
Ocular findings in a family with autosomal dominant retinitis pigmentosa and a frameshift mutation altering the carboxyl terminal sequence of rhodopsin.
1993,
Pubmed
Baehr,
Characterization of bovine rod outer segment G-protein.
1982,
Pubmed
Berbari,
Bardet-Biedl syndrome proteins are required for the localization of G protein-coupled receptors to primary cilia.
2008,
Pubmed
Bessant,
Severe autosomal dominant retinitis pigmentosa caused by a novel rhodopsin mutation (Ter349Glu). Mutations in brief no. 208. Online.
1999,
Pubmed
Chan,
Knock-in human rhodopsin-GFP fusions as mouse models for human disease and targets for gene therapy.
2004,
Pubmed
Chiang,
Comparative genomic analysis identifies an ADP-ribosylation factor-like gene as the cause of Bardet-Biedl syndrome (BBS3).
2004,
Pubmed
Chuang,
The cytoplasmic tail of rhodopsin acts as a novel apical sorting signal in polarized MDCK cells.
1998,
Pubmed
Concepcion,
Q344ter mutation causes mislocalization of rhodopsin molecules that are catalytically active: a mouse model of Q344ter-induced retinal degeneration.
2010,
Pubmed
Davis,
A knockin mouse model of the Bardet-Biedl syndrome 1 M390R mutation has cilia defects, ventriculomegaly, retinopathy, and obesity.
2007,
Pubmed
Deretic,
Regulation of sorting and post-Golgi trafficking of rhodopsin by its C-terminal sequence QVS(A)PA.
1998,
Pubmed
Gross,
Defective development of photoreceptor membranes in a mouse model of recessive retinal degeneration.
2006,
Pubmed
Hagstrom,
Retinal degeneration in tulp1-/- mice: vesicular accumulation in the interphotoreceptor matrix.
1999,
Pubmed
Hagstrom,
Recessive mutations in the gene encoding the tubby-like protein TULP1 in patients with retinitis pigmentosa.
1998,
Pubmed
Hashimoto,
Lentiviral gene replacement therapy of retinas in a mouse model for Usher syndrome type 1B.
2007,
Pubmed
Hollingsworth,
Defective trafficking of rhodopsin and its role in retinal degenerations.
2012,
Pubmed
Illing,
A rhodopsin mutant linked to autosomal dominant retinitis pigmentosa is prone to aggregate and interacts with the ubiquitin proteasome system.
2002,
Pubmed
Jin,
The conserved Bardet-Biedl syndrome proteins assemble a coat that traffics membrane proteins to cilia.
2010,
Pubmed
Lakso,
Efficient in vivo manipulation of mouse genomic sequences at the zygote stage.
1996,
Pubmed
Lehtokari,
Identification of a founder mutation in TPM3 in nemaline myopathy patients of Turkish origin.
2008,
Pubmed
Lem,
Morphological, physiological, and biochemical changes in rhodopsin knockout mice.
1999,
Pubmed
Leroux,
Prenatal diagnosis of recessive congenital methaemoglobinaemia type II: novel mutation in the NADH-cytochrome b5 reductase gene leading to stop codon read-through.
2005,
Pubmed
Liang,
Rhodopsin signaling and organization in heterozygote rhodopsin knockout mice.
2004,
Pubmed
Mazelova,
Ciliary targeting motif VxPx directs assembly of a trafficking module through Arf4.
2009,
Pubmed
,
Xenbase
Melia,
A comparison of the efficiency of G protein activation by ligand-free and light-activated forms of rhodopsin.
1997,
Pubmed
Nachury,
A core complex of BBS proteins cooperates with the GTPase Rab8 to promote ciliary membrane biogenesis.
2007,
Pubmed
Nakao,
The role of mislocalized phototransduction in photoreceptor cell death of retinitis pigmentosa.
2012,
Pubmed
Nishimura,
Bbs2-null mice have neurosensory deficits, a defect in social dominance, and retinopathy associated with mislocalization of rhodopsin.
2004,
Pubmed
Peters,
Fine structure of a periciliary ridge complex of frog retinal rod cells revealed by ultrahigh resolution scanning electron microscopy.
1983,
Pubmed
Portera-Cailliau,
Apoptotic photoreceptor cell death in mouse models of retinitis pigmentosa.
1994,
Pubmed
Price,
Mislocalization and degradation of human P23H-rhodopsin-GFP in a knockin mouse model of retinitis pigmentosa.
2011,
Pubmed
Robinson,
Assays for detection of constitutively active opsins.
2000,
Pubmed
Sancho-Pelluz,
Mice with a D190N mutation in the gene encoding rhodopsin: a model for human autosomal-dominant retinitis pigmentosa.
2012,
Pubmed
Schlüter,
Trafficking of Crumbs3 during cytokinesis is crucial for lumen formation.
2009,
Pubmed
Seo,
BBS6, BBS10, and BBS12 form a complex with CCT/TRiC family chaperonins and mediate BBSome assembly.
2010,
Pubmed
Sung,
A rhodopsin gene mutation responsible for autosomal dominant retinitis pigmentosa results in a protein that is defective in localization to the photoreceptor outer segment.
1994,
Pubmed
Swiderski,
Gene expression analysis of photoreceptor cell loss in bbs4-knockout mice reveals an early stress gene response and photoreceptor cell damage.
2007,
Pubmed
Tam,
The role of rhodopsin glycosylation in protein folding, trafficking, and light-sensitive retinal degeneration.
2009,
Pubmed
,
Xenbase
Zeitz,
Identification and functional characterization of a novel rhodopsin mutation associated with autosomal dominant CSNB.
2008,
Pubmed
Zirn,
Novel familial WT1 read-through mutation associated with Wilms tumor and slow progressive nephropathy.
2005,
Pubmed
