Mutations in the gene encoding the disk rim specific protein, peripherin/rds, have been implicated in the pathogenesis of both autosomal dominant retinitis pigmentosa (ADRP) and various forms of macular dystrophy (MD). Peripherin/rds plays an important role in the morphogenesis and maintenance of the disk rim structure. This role is supported by interactions with other proteins. The primary goals of the proposed research are to determine the molecular basis of action and functional properties of peripherin/rds and these interacting proteins, and to investigate the molecular abnormalities that lead to photoreceptor degeneration in transgenic models of human retinal diseases.
Specific Aims 1, 2 and 3 are to use the structural, electrophysiological and biochemical studies to evaluate transgenic retinas expressing different mutations in the peripherin/rds gene. These mutations include: (1) the ARG172TP mutation that causes MD in humans, (2) the CYS214Ser mutation to determine the role of intermolecular disulfide bonds in the assembly of functional peripherin/rds-rom-1 complexes. We will analyze the effects of these mutations on morphogenesis of the disc membrane and on regulation of complex formation by peripherin/rds and rom-1. We can evaluate the different roles played by peripherin/rds in rods versus cones by comparing the effects of the Arg172Trp and Cys214Ser mutations on the structure and function of rod and cone outer segments. Comparisons of the Cys214 and Cys150Ser transgenic mice will provide insight into the role of inter- and intramolecular disulfide bond formation in peripherin/rds function. In Specific im 4, we will use a yeast two-hybrid system to identify the sites of interaction involved in formation of multimeric complexes by peripherin/rds and rom-1. This system provides a powerful genetic mechanism for detecting protein-protein interactions. We will also use the yeast two-hybrid system to screen a retinal cDNA library for other proteins that are involved in the assembly of functional peripherin/rds-rom-1 complexes. We have identified regions of high homology between all known peripherin/rds and rom-1 that are located in the large intradiscal loop. We hypothesize that these regions mediate the interactions between peripherin/rds-rom-1 complexes and are required to hold the rod discs or cone lamellae in their flattened shape. These interactions may be mediated by direct association of the subunit complexes or indirectly through other proteins. Mutagenesis studies will be performed to evaluate these interactions. These studies will provide insight into the functional role of peripherin/rds in normal and diseased retinas.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY010609-05
Application #
6164681
Study Section
Visual Sciences C Study Section (VISC)
Program Officer
Dudley, Peter A
Project Start
1995-08-01
Project End
2000-08-31
Budget Start
2000-03-01
Budget End
2000-08-31
Support Year
5
Fiscal Year
2000
Total Cost
$203,587
Indirect Cost
Name
University of Illinois at Chicago
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Kelley, Ryan A; Al-Ubaidi, Muayyad R; Naash, Muna I (2018) Retbindin Is Capable of Protecting Photoreceptors from Flavin-Sensitized Light-Mediated Cell Death In Vitro. Adv Exp Med Biol 1074:485-490
LaVail, Matthew M; Nishikawa, Shimpei; Steinberg, Roy H et al. (2018) Phenotypic characterization of P23H and S334ter rhodopsin transgenic rat models of inherited retinal degeneration. Exp Eye Res 167:56-90
Ikelle, Larissa; Naash, Muna I; Al-Ubaidi, Muayyad R (2018) Role of Fibulins 2 and 5 in Retinal Development and Maintenance. Adv Exp Med Biol 1074:275-280
Zulliger, Rahel; Conley, Shannon M; Mwoyosvi, Maggie L et al. (2018) Oligomerization of Prph2 and Rom1 is essential for photoreceptor outer segment formation. Hum Mol Genet 27:3507-3518
Sinha, Tirthankar; Makia, Mustafa; Du, Jianhai et al. (2018) Flavin homeostasis in the mouse retina during aging and degeneration. J Nutr Biochem 62:123-133
Agbaga, Martin-Paul; Merriman, Dana K; Brush, Richard S et al. (2018) Differential composition of DHA and very-long-chain PUFAs in rod and cone photoreceptors. J Lipid Res 59:1586-1596
Conley, Shannon M; Stuck, Michael W; Watson, Jamie N et al. (2017) Rom1 converts Y141C-Prph2-associated pattern dystrophy to retinitis pigmentosa. Hum Mol Genet 26:509-518
Kelley, Ryan A; Al-Ubaidi, Muayyad R; Sinha, Tirthankar et al. (2017) Ablation of the riboflavin-binding protein retbindin reduces flavin levels and leads to progressive and dose-dependent degeneration of rods and cones. J Biol Chem 292:21023-21034
Stuck, Michael W; Conley, Shannon M; Naash, Muna I (2016) PRPH2/RDS and ROM-1: Historical context, current views and future considerations. Prog Retin Eye Res 52:47-63
Kelley, Ryan A; Al-Ubaidi, Muayyad R; Naash, Muna I (2016) The Potential Role of Flavins and Retbindin in Retinal Function and Homeostasis. Adv Exp Med Biol 854:643-8

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