Document Detail

Localization of ZO-1 in the nucleolus of corneal fibroblasts.
MedLine Citation:
PMID:  17460259     Owner:  NLM     Status:  MEDLINE    
PURPOSE: Within the multidomain structure of ZO-1 are motifs responsible for ZO-1's localization to intercellular junctions and its newly demonstrated localization to the leading edge of lamellipodia in corneal fibroblasts. Since ZO-1 also has two nuclear localization signals, this study was undertaken to determine whether stimuli associated with wounding would induce nuclear translocation of ZO-1 METHODS: Immunocytochemistry and immunoblot analysis were used to localize endogenous and exogenous ZO-1 in nuclear and cytoplasmic sites in corneal fibroblasts and 293T fibroblasts, with and without myc-ZO-1 transfection. Cells were serum starved by growth for 48 hours in DMEM/F12 with 0.2% FBS and subsequently were either scrape wounded or treated with 10% FBS, PDGF, or FGF-2 for 6 hours. For immunoblot analysis, after lysis, the nuclear and cytosolic fractions were separated and analyzed by SDS-PAGE. Cells on companion coverslips were fixed with 3% p-formaldehyde and permeabilized with 1% Triton before immunocytochemical detection of ZO-1 and nuclear proteins. RESULTS: ZO-1 was rarely detected in the nucleus of serum-starved corneal fibroblasts. In contrast, it colocalized with nucleolin in the nucleoli of corneal fibroblasts after serum-starved cells were treated with 10% FBS, PDGF, or FGF-2. Immunoblot analysis confirmed the immunocytochemical results: Little ZO-1 was detected in the nuclear fraction of lysates of serum-starved cells, but ZO-1 was found in the nuclear fractions of rabbit corneal and 293T fibroblasts treated with 10% FBS, PDGF, or FGF-2. Furthermore in scrape-wounded corneal fibroblasts, ZO-1 was localized to nucleoli of both serum-starved and serum-treated cells. CONCLUSIONS: Localization of ZO-1 to nucleoli of corneal and 293T fibroblasts under proliferative and promigratory conditions suggests a physiologically significant interaction of ZO-1 with proteins in nucleoli during the healing process.
Miriam Benezra; Roseanne S Greenberg; Sandra K Masur
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  48     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-04-26     Completed Date:  2007-06-13     Revised Date:  2007-12-03    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2043-9     Citation Subset:  IM    
Department of Ophthalmology, Mount Sinai School of Medicine, New York, New York 10029-6574, USA.
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MeSH Terms
Cell Line
Cell Movement / physiology
Cell Nucleolus / metabolism*
Cornea / injuries,  metabolism*
Electrophoresis, Polyacrylamide Gel
Fibroblasts / metabolism
Fluorescent Antibody Technique, Indirect
Membrane Proteins / genetics,  metabolism*
Phosphoproteins / genetics,  metabolism*
RNA-Binding Proteins / metabolism
Wound Healing / physiology
Grant Support
1 R24 CA095823/CA/NCI NIH HHS; P30 EY001867/EY/NEI NIH HHS; R01-EY09414/EY/NEI NIH HHS
Reg. No./Substance:
0/Membrane Proteins; 0/Phosphoproteins; 0/RNA-Binding Proteins; 0/nucleolin; 0/zonula occludens-1 protein

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