MOLECULAR ANALYSIS OF OCULAR LENS BEADED FILAMENTS

Summary

Principal Investigator: Paul Fitzgerald
Affiliation: University of California
Country: USA
Abstract: We seek to understand the function of Intermediate Filaments in the ocular lens, with a special emphasis on the Beaded Filament, a derivative of the Intermediate Filament which is unique to the lens fiber cell. To accomplish this we propose studies which will: 1. Identify Beaded Filament- and Intermediate Filament Associated Proteins. Identification of such proteins will yield clues to the function of these filaments in the lens, and lead to an understanding of how these filaments integrate into lens cell biology. 2. Identify structural relationships between Beaded/Intermediate Filaments and the lens cell. Identification of structural relationships will again suggest hypotheses of function, and delineate how the filaments are integrated into lens cell biology. 3. Define the molecular relationships of beaded filament proteins in the intact beaded filament, so that the impact of cataract-causing mutations can be understood. 4. Define the dynamics of Beaded/Intermediate Filaments in living lens cells, to gain an understanding of what functions these structures play in the lens. These studies will utilize a range of cell biological and molecular techniques, and take advantage of opportunities provided by knockout mice lacking CP49, CP95, and vimentin. We will employ light and electron microscopy, immunochemical approaches, molecular cloning, protein purification and identification, site-directed spin labeling/electron paramagnetic resonance, confocal microscopy, transfection, and transgenic approaches. Completion of these studies will explain the function of theses proteins/structures in normal lens biology, permitting us to understand how inherited human mutations in these proteins result in cataract.
Funding Period: 1990-08-01 - 2008-07-31
more information: NIH RePORT

Top Publications

  1. pmc Characterization of structural changes in vimentin bearing an epidermolysis bullosa simplex-like mutation using site-directed spin labeling and electron paramagnetic resonance
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    J Biol Chem 280:2141-6. 2005
  2. pmc Characterization of the linker 2 region in human vimentin using site-directed spin labeling and electron paramagnetic resonance
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Biochemistry 45:11737-43. 2006
  3. pmc Treatment of keratin intermediate filaments with sulfur mustard analogs
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, 1 Shields Ave, Davis, CA 95616 8643, USA
    Biochem Biophys Res Commun 359:616-21. 2007
  4. ncbi Structural specializations emerging late in mouse lens fiber cell differentiation
    Tom Blankenship
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Invest Ophthalmol Vis Sci 48:3269-76. 2007
  5. pmc Identifying the role of specific motifs in the lens fiber cell specific intermediate filament phakosin
    Joshua T Pittenger
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, CA 95616, USA
    Invest Ophthalmol Vis Sci 48:5132-41. 2007
  6. doi Resisting the effects of aging: a function for the fiber cell beaded filament
    Kyoung hye Yoon
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Invest Ophthalmol Vis Sci 49:1030-6. 2008
  7. pmc Identification of phosphorylation-induced changes in vimentin intermediate filaments by site-directed spin labeling and electron paramagnetic resonance
    Josh T Pittenger
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Biochemistry 47:10863-70. 2008
  8. pmc Periplakin interactions with lens intermediate and beaded filaments
    Kyoung hye Yoon
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California, USA
    Invest Ophthalmol Vis Sci 50:1283-9. 2009
  9. pmc Lens intermediate filaments
    Paul G FitzGerald
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, CA 95616, USA
    Exp Eye Res 88:165-72. 2009

Scientific Experts

  • J F Hess
  • Paul Fitzgerald
  • Kyoung hye Yoon
  • Tom Blankenship
  • Josh T Pittenger
  • Bradley Shibata
  • Joshua T Pittenger
  • John C Voss
  • Madhu S Budamagunta
  • Linsey Bradshaw

Detail Information

Publications9

  1. pmc Characterization of structural changes in vimentin bearing an epidermolysis bullosa simplex-like mutation using site-directed spin labeling and electron paramagnetic resonance
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    J Biol Chem 280:2141-6. 2005
    ..These data point toward a specific stage of assembly at which a common disease-causing mutation in IF proteins interrupts assembly...
  2. pmc Characterization of the linker 2 region in human vimentin using site-directed spin labeling and electron paramagnetic resonance
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Biochemistry 45:11737-43. 2006
    ..In conjunction with our previous characterization of region 305-336 and the solved structure of rod 2B from 328-405, the full extent of coiled-coil domain in rod 2B is now known, spanning from vimentin positions 302-405...
  3. pmc Treatment of keratin intermediate filaments with sulfur mustard analogs
    John F Hess
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, 1 Shields Ave, Davis, CA 95616 8643, USA
    Biochem Biophys Res Commun 359:616-21. 2007
    ..Our results are consistent with the hypothesis that modification of keratins by SM would result in keratin filament destruction, leading to lysis of epidermal basal cells and skin blistering...
  4. ncbi Structural specializations emerging late in mouse lens fiber cell differentiation
    Tom Blankenship
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Invest Ophthalmol Vis Sci 48:3269-76. 2007
    ..To describe a previously uncharacterized structural specialization in the mouse lens fiber cell and to delineate its emergence relative to lens development and fiber cell differentiation...
  5. pmc Identifying the role of specific motifs in the lens fiber cell specific intermediate filament phakosin
    Joshua T Pittenger
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, CA 95616, USA
    Invest Ophthalmol Vis Sci 48:5132-41. 2007
    ..In this report we test specific motifs/domains in phakosin to identify changes that that have adapted phakosin to lens-specific structure and function...
  6. doi Resisting the effects of aging: a function for the fiber cell beaded filament
    Kyoung hye Yoon
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Invest Ophthalmol Vis Sci 49:1030-6. 2008
    ..It includes phakosin and filensin, two divergent members of the intermediate filament family of proteins that are also unique to the fiber cell. The authors sought to determine what function the beaded filament fulfills in the lens...
  7. pmc Identification of phosphorylation-induced changes in vimentin intermediate filaments by site-directed spin labeling and electron paramagnetic resonance
    Josh T Pittenger
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA
    Biochemistry 47:10863-70. 2008
    ....
  8. pmc Periplakin interactions with lens intermediate and beaded filaments
    Kyoung hye Yoon
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California, USA
    Invest Ophthalmol Vis Sci 50:1283-9. 2009
    ..The goal of this study was to identify linker proteins that integrate the different lens IF into the biology of the lens fiber cells...
  9. pmc Lens intermediate filaments
    Paul G FitzGerald
    Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, CA 95616, USA
    Exp Eye Res 88:165-72. 2009
    ..Some of the key features of both filament systems are reviewed...