期刊
HUMAN MOLECULAR GENETICS
卷 29, 期 12, 页码 2076-2097出版社
OXFORD UNIV PRESS
DOI: 10.1093/hmg/ddaa096
关键词
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资金
- National Institutes of Health/National Eye Institute [R01 EY021505, R01 EY029770, R21 EY027389]
- Knights Templar Pediatric Ophthalmology Career Starter Grant Award
- University of Delaware Research Foundation (UDRF), Inc.
- UDRFStrategic Initiatives Award
- National Science Foundation Fellowship from the Greater Philadelphia Region Louis Stokes Alliance for Minority Participation (LSAMP) Bridge to the Doctorate (BTD) Program
- University of Delaware
- Fight For Sight Summer Student Fellowship
- University of Delaware Center for Bioinformatics and Computational Biology Core Facility, Core Imaging facility, Sequencing and Genotyping Center
- National Institutes of Health/National Institute of General Medical Sciences INBRE ProgramGrant [P20 GM103446]
- National Institutes of Health/National Center for Research Resources grant [1S10 RR027273-01]
- National Institutes of Health/National Institute of General Medical Sciences [P30 GM114736]
- State of Delaware
Mutations of the RNA granule component TDRD7 (OMIM: 611258) cause pediatric cataract. We applied an integrated approach to uncover the molecular pathology of cataract in Tdrd7-/- mice. Early postnatal Tdrd7-/- animals precipitously develop cataract suggesting a global-level breakdown/misregulation of key cellular processes. High-throughput RNA sequencing integrated with iSyTE-bioinformatics analysis identified the molecular chaperone and cytoskeletal modulator, HSPB1, among high-priority downregulated candidates in Tdrd7-/- lens. A protein fluorescence two-dimensional difference in-gel electrophoresis (2D-DIGE)-coupled mass spectrometry screen also identified HSPB1 downregulation, offering independent support for its importance to Tdrd7-/- cataractogenesis. Lens fiber cells normally undergo nuclear degradation for transparency, posing a challenge: how is their cell morphology, also critical for transparency, controlled post-nuclear degradation? HSPB1 functions in cytoskeletal maintenance, and its reduction in Tdrd7-/- lens precedes cataract, suggesting cytoskeletal defects may contribute to Tdrd7-/- cataract. In agreement, scanning electron microscopy (SEM) revealed abnormal fiber cell morphology in Tdrd7-/- lenses. Further, abnormal phalloidin and wheat germ agglutinin (WGA) staining of Tdrd7-/- fiber cells, particularly those exhibiting nuclear degradation, reveals distinct regulatory mechanisms control F-actin cytoskeletal and/or membrane maintenance in post-organelle degradation maturation stage fiber cells. Indeed, RNA immunoprecipitation identified Hspb1 mRNA in wild-type lens lysate TDRD7-pulldowns, and single-molecule RNA imaging showed co-localization of TDRD7 protein with cytoplasmic Hspb1 mRNA in differentiating fiber cells, suggesting that TDRD7-ribonucleoprotein complexes may be involved in optimal buildup of key factors. Finally, Hspb1 knockdown in Xenopus causes eye/lens defects. Together, these data uncover TDRD7's novel upstream role in elevation of stress-responsive chaperones for cytoskeletal maintenance in post-nuclear degradation lens fiber cells, perturbation of which causes early-onset cataracts.
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