| Localization of ZO-1 in the nucleolus of corneal fibroblasts. | |
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MedLine Citation:
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PMID: 17460259 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Miriam Benezra; Roseanne S Greenberg; Sandra K Masur |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Investigative ophthalmology & visual science Volume: 48 ISSN: 0146-0404 ISO Abbreviation: Invest. Ophthalmol. Vis. Sci. Publication Date: 2007 May |
Date Detail:
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Created Date: 2007-04-26 Completed Date: 2007-06-13 Revised Date: 2007-12-03 |
Medline Journal Info:
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Nlm Unique ID: 7703701 Medline TA: Invest Ophthalmol Vis Sci Country: United States |
Other Details:
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Languages: eng Pagination: 2043-9 Citation Subset: IM |
Affiliation:
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Department of Ophthalmology, Mount Sinai School of Medicine, New York, New York 10029-6574, USA. benezram@mskcc.org |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Cell Movement / physiology Cell Nucleolus / metabolism* Cornea / injuries, metabolism* Cytosol Electrophoresis, Polyacrylamide Gel Fibroblasts / metabolism Fluorescent Antibody Technique, Indirect Immunoblotting Membrane Proteins / genetics, metabolism* Phosphoproteins / genetics, metabolism* RNA-Binding Proteins / metabolism Rabbits Transfection Wound Healing / physiology |
| Grant Support | |
ID/Acronym/Agency:
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1 R24 CA095823/CA/NCI NIH HHS; P30 EY001867/EY/NEI NIH HHS; R01-EY09414/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Membrane Proteins; 0/Phosphoproteins; 0/RNA-Binding Proteins; 0/nucleolin; 0/zonula occludens-1 protein |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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