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 Myo/Nog cells: targets for preventing the accumulation of skeletal muscle-like cells in the human lens
    Jacquelyn Gerhart
    Lankenau Institute for Medical Research, Wynnewood, Pennsylvania, United States of America
    PLoS ONE 9:e95262. 2014
  2. pmc Carbon turnover in the water-soluble protein of the adult human lens
    Daniel N Stewart
    Department of Chemistry, University of California, One Shields Avenue, Davis, CA, USA
    Mol Vis 19:463-75. 2013
  3. pmc A role for γS-crystallin in the organization of actin and fiber cell maturation in the mouse lens
    Jianguo Fan
    Section on Molecular Structure and Functional Genomics, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 0608, USA
    FEBS J 279:2892-904. 2012
  4. pmc Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane
    John J Castorino
    Department of Pathology, Anatomy, and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    Invest Ophthalmol Vis Sci 52:6774-84. 2011
  5. pmc Further analysis of the lens phenotype in Lim2-deficient mice
    Yanrong Shi
    Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri 63110, USA
    Invest Ophthalmol Vis Sci 52:7332-9. 2011
  6. pmc Intermediate filaments regulate tissue size and stiffness in the murine lens
    Douglas S Fudge
    Department of Integrative Biology, University of Guelph, Guelph, Ontario, Canada
    Invest Ophthalmol Vis Sci 52:3860-7. 2011
  7. 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
  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 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
  10. 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

Scientific Experts

  • J F Hess
  • Paul Fitzgerald
  • Kyoung hye Yoon
  • Tom Blankenship
  • Jacquelyn Gerhart
  • Daniel N Stewart
  • Jianguo Fan
  • Douglas S Fudge
  • Huan Wang
  • Yanrong Shi
  • Richard T Mathias
  • John J Castorino
  • Josh T Pittenger
  • Bradley Shibata
  • Joshua T Pittenger
  • Arturo Bravo-Nuevo
  • Meghan Pitts
  • Marvin Greenbaum
  • Mindy George-Weinstein
  • Mitchell Crawford
  • Victoria Scheinfeld
  • Miranda J S Falso
  • Bruce D Hammock
  • Bruce A Buchholz
  • Krishnan P Nambiar
  • David M Rocke
  • Jozsef Lango
  • Lijin Dong
  • Sanghamitra Mishra
  • Yingwei Chen
  • Graeme Wistow
  • Jessica Polishook
  • Barbara Kloeckener-Gruissem
  • Alex V Levin
  • Alicia B De Maria
  • Shannon Van Stralen
  • John V McCuaig
  • Shannon M Gallagher-Colombo
  • Eric Ostertag
  • Nancy J Philp
  • Steven Bassnett
  • John C Voss
  • Madhu S Budamagunta
  • Linsey Bradshaw

Detail Information

Publications15

  1. pmc Myo/Nog cells: targets for preventing the accumulation of skeletal muscle-like cells in the human lens
    Jacquelyn Gerhart
    Lankenau Institute for Medical Research, Wynnewood, Pennsylvania, United States of America
    PLoS ONE 9:e95262. 2014
    ..Myo/Nog cells appear to be the source of skeletal muscle-like cells in explants of human lens tissue. Targeting Myo/Nog cells with the G8 antibody during cataract surgery may reduce the incidence of PCO...
  2. pmc Carbon turnover in the water-soluble protein of the adult human lens
    Daniel N Stewart
    Department of Chemistry, University of California, One Shields Avenue, Davis, CA, USA
    Mol Vis 19:463-75. 2013
    ....
  3. pmc A role for γS-crystallin in the organization of actin and fiber cell maturation in the mouse lens
    Jianguo Fan
    Section on Molecular Structure and Functional Genomics, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 0608, USA
    FEBS J 279:2892-904. 2012
    ..This suggests that γS may have a functional role related to actin, perhaps in 'shepherding' filaments to maintain the optical properties of the lens cytoplasm and normal fiber cell maturation...
  4. pmc Juvenile cataract-associated mutation of solute carrier SLC16A12 impairs trafficking of the protein to the plasma membrane
    John J Castorino
    Department of Pathology, Anatomy, and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    Invest Ophthalmol Vis Sci 52:6774-84. 2011
    ..Q215X) causes juvenile cataract with a dominant inheritance pattern. In the present study, in vitro and in vivo experimental models were used to gain insight into how the SLC16A12 (c.643C>T) mutation leads to cataract formation...
  5. pmc Further analysis of the lens phenotype in Lim2-deficient mice
    Yanrong Shi
    Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri 63110, USA
    Invest Ophthalmol Vis Sci 52:7332-9. 2011
    ..A comparative analysis was performed of wild-type and Lim2-deficient (Lim2(Gt/Gt)) mouse lenses, to better define the anatomic and physiologic roles of Lim2...
  6. pmc Intermediate filaments regulate tissue size and stiffness in the murine lens
    Douglas S Fudge
    Department of Integrative Biology, University of Guelph, Guelph, Ontario, Canada
    Invest Ophthalmol Vis Sci 52:3860-7. 2011
    ..To define the contributions of the beaded filament (BF), a lens-specific intermediate filament (IF), to lens morphology and biomechanics...
  7. 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...
  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 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
    ....
  10. 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...
  11. 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...
  12. 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...
  13. 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...
  14. 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...
  15. 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...