Molecular Mechanisms in Retinal Degeneration

Summary

Principal Investigator: Debora Farber
Affiliation: University of California
Country: USA
Abstract: DESCRIPTION (provided by applicant): The long-term objectives of this proposal are to characterize genes involved in human retinal degenerations and to find possible ways to halt or eventually cure these blinding diseases. Specifically, we will study two proteins present in cones, 15A15 and retinoschisin, encoded by genes isolated previously in our laboratory, that may be interrelated in their function. We have found that the 15A15 protein binds to several DNA fragments containing the core consensus sequences for hormone response elements and that it has several features characteristic of coactivators/ co-repressors of nuclear hormone receptors (NHRs). In our first Specific Aim we will: corroborate that 15A15 plays this role studying its protein-protein interactions with GST pull-down assays;investigate if 15A15 interactions with NHRs are hormone-dependent and enhance the transactivation of endogenous NHRs;map the region of 15A15 necessary for these interactions;determine if 15A15 binds directly to response elements present in the promoter of retinoschisin and other genes expressed in cones, regulating their transcription, and if mutations in 15A15 modules abolish transcriptional enhancement of NHRs. Chromatin immunoprecipitation assays will show if 15A15 exists as part of a complex with other factors, in vivo. We will also screen the DNA of patients with retinal degenerations affecting cones for mutations in 15A15 that may result in disease. In our second Specific Aim, we will continue studying retinoschisin, the secreted protein involved in cell-cell interactions that when mutated causes X-linked juvenile retinoschisis (XLRS). We have found that cGMP, Ca2+ and possibly G-proteins may participate in regulation of secretion of retinoschisin from the photoreceptor inner segments. On the basis of these data, we will carry out a systematic study on the mechanisms that control this process, investigating the involvement of membrane and soluble guanylate cyclases (GC), the effects of GC-activating proteins, nitric oxide and carbon monoxide;the participation of rod, cone and Ca2+/calmodulin-dependent PDEs, and the role of guanine deaminase, which converts cGMP into cXMP. We will also determine whether L-type voltage gated Ca2+ channels and intracellular Ca2+ stores, as well as Gbeta-gamma and mGluR8 influence the secretion of retinoschisin. For each photoreceptor component studied, we will start with a pharmacological approach to establish its involvement, followed by its removal from the system with the use of shRNAs to corroborate its effect. Overall, our research will increase the understanding of the interaction and function of 15A15 and retinoschisin, two important proteins in cones. By learning about the biochemical pathways involved in the regulation of retinoschisin secretion in normal physiological conditions, that may be important for potential pharmacological treatment of XLRS, we hope to open new avenues to explore possible ways to rescue vision for diseases for which there are no current cures.
Funding Period: 1989-08-01 - 2011-11-30
more information: NIH RePORT

Top Publications

  1. pmc Sequence-specific binding of recombinant Zbed4 to DNA: insights into Zbed4 participation in gene transcription and its association with other proteins
    Vladislav V Mokhonov
    Jules Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America
    PLoS ONE 7:e35317. 2012
  2. pmc Dynamics of the rhomboid-like protein RHBDD2 expression in mouse retina and involvement of its human ortholog in retinitis pigmentosa
    Novruz B Ahmedli
    Jules Stein Eye Institute, UCLA, Los Angeles, California 90095, USA
    J Biol Chem 288:9742-54. 2013
  3. pmc Regulation of retinoschisin secretion in Weri-Rb1 cells by the F-actin and microtubule cytoskeleton
    Eiko Kitamura
    Jules Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America
    PLoS ONE 6:e20707. 2011
  4. pmc ZBED4, a cone and Müller cell protein in human retina, has a different cellular expression in mouse
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, UCLA, Los Angeles, CA, USA
    Mol Vis 17:2011-8. 2011
  5. ncbi Two forms of the large tumor suppressor gene (Lats1) protein expressed in the vertebrate retina
    Novrouz B Akhmedov
    Jules Stein Eye Institute, UCLA School of Medicine, 100 Stein Plaza, Los Angeles, CA 90095, USA
    Biochim Biophys Acta 1728:11-7. 2005
  6. ncbi Disruption of the gene encoding the beta1-subunit of transducin in the Rd4/+ mouse
    Eiko Kitamura
    Jules Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA
    Invest Ophthalmol Vis Sci 47:1293-301. 2006
  7. ncbi Replication and refinement of linkage of posterior polymorphous corneal dystrophy to the posterior polymorphous corneal dystrophy 1 locus on chromosome 20
    Vivek S Yellore
    Cornea Service, The Jules Stein Eye Institute, Department of Human Genetics, and Retinal Biochemistry Laboratory, David Geffen School of Medicine at University of California Los Angeles, California 90095, USA
    Genet Med 9:228-34. 2007
  8. doi Identification and characterization of genes expressed in cone photoreceptors
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, David Geffen School of Medicine and Molecular Biology Institute, UCLA, Los Angeles, California 90095, USA
    Adv Exp Med Biol 613:235-44. 2008
  9. pmc ZBED4, a BED-type zinc-finger protein in the cones of the human retina
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, David Geffen School of Medicine, University of California Los Angeles UCLA, Los Angeles, California, USA
    Invest Ophthalmol Vis Sci 50:3580-8. 2009

Scientific Experts

  • Debora Farber
  • Novrouz B Akhmedov
  • Mehrnoosh Saghizadeh
  • Eiko Kitamura
  • Novruz B Ahmedli
  • Yekaterina Gribanova
  • Vladislav V Mokhonov
  • Yekaterina E Gribanova
  • Emmanuel Mendoza
  • Clyde K Yamashita
  • Vivek S Yellore
  • Riza Koksal Ozgul
  • Jerry E Johnson
  • Collins C Njoku
  • Donald A Fox
  • Akash Naidu
  • Alejandra Young
  • Veena P Theendakara
  • Veena Theendakara
  • Alexander V Ljubimov
  • Stanley F Nelson
  • Sylvia A Rayner
  • M Ali Khan
  • Eric Sobel
  • Jeanette C Papp
  • Anthony J Aldave
  • Steven Nusinowitz
  • Bo Chang
  • Nagesh P Rao
  • Michael Danciger
  • Clyde Yamashita

Detail Information

Publications9

  1. pmc Sequence-specific binding of recombinant Zbed4 to DNA: insights into Zbed4 participation in gene transcription and its association with other proteins
    Vladislav V Mokhonov
    Jules Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America
    PLoS ONE 7:e35317. 2012
    ..Most importantly, in vitro experiments using constructs containing promoters of genes directing expression of the luciferase gene, showed that Zbed4 transactivates the transcription of those promoters with poly-G tracts...
  2. pmc Dynamics of the rhomboid-like protein RHBDD2 expression in mouse retina and involvement of its human ortholog in retinitis pigmentosa
    Novruz B Ahmedli
    Jules Stein Eye Institute, UCLA, Los Angeles, California 90095, USA
    J Biol Chem 288:9742-54. 2013
    ..Our findings suggest that the RHBDD2 protein plays important roles in the development and normal function of the retina...
  3. pmc Regulation of retinoschisin secretion in Weri-Rb1 cells by the F-actin and microtubule cytoskeleton
    Eiko Kitamura
    Jules Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America
    PLoS ONE 6:e20707. 2011
    ....
  4. pmc ZBED4, a cone and Müller cell protein in human retina, has a different cellular expression in mouse
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, UCLA, Los Angeles, CA, USA
    Mol Vis 17:2011-8. 2011
    ..To begin investigating these potential roles of ZBED4, we studied the developmental expression and localization of both the Zbed4 mRNA and protein of mouse retina...
  5. ncbi Two forms of the large tumor suppressor gene (Lats1) protein expressed in the vertebrate retina
    Novrouz B Akhmedov
    Jules Stein Eye Institute, UCLA School of Medicine, 100 Stein Plaza, Los Angeles, CA 90095, USA
    Biochim Biophys Acta 1728:11-7. 2005
    ..The expression of both isoforms in vertebrate cells raises the possibility that these Lats1 proteins may act as negative key regulators of the cell cycle, each of them performing a unique role...
  6. ncbi Disruption of the gene encoding the beta1-subunit of transducin in the Rd4/+ mouse
    Eiko Kitamura
    Jules Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA
    Invest Ophthalmol Vis Sci 47:1293-301. 2006
    ..The hypothesis for the study was that disruption of a gene at one of the two breakpoints in the Rd4 chromosome is responsible for the retinal degeneration. The purpose was to identify the disrupted gene...
  7. ncbi Replication and refinement of linkage of posterior polymorphous corneal dystrophy to the posterior polymorphous corneal dystrophy 1 locus on chromosome 20
    Vivek S Yellore
    Cornea Service, The Jules Stein Eye Institute, Department of Human Genetics, and Retinal Biochemistry Laboratory, David Geffen School of Medicine at University of California Los Angeles, California 90095, USA
    Genet Med 9:228-34. 2007
    ..Glaucoma also develops in up to 40% of patients with posterior polymorphous corneal dystrophy...
  8. doi Identification and characterization of genes expressed in cone photoreceptors
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, David Geffen School of Medicine and Molecular Biology Institute, UCLA, Los Angeles, California 90095, USA
    Adv Exp Med Biol 613:235-44. 2008
  9. pmc ZBED4, a BED-type zinc-finger protein in the cones of the human retina
    Mehrnoosh Saghizadeh
    Jules Stein Eye Institute, David Geffen School of Medicine, University of California Los Angeles UCLA, Los Angeles, California, USA
    Invest Ophthalmol Vis Sci 50:3580-8. 2009
    ..To characterize the ZBED4 cDNA identified by subtractive hybridization and microarray of retinal cone degeneration (cd) adult dog mRNA from mRNA of normal dog retina...