| Nuclear matrix-bound replicational sites detected in situ by 5-bromodeoxyuridine. | |
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MedLine Citation:
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PMID: 1429014 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The nuclear matrix was prepared in situ from Swiss 3T3 cells, which were synchronized by contact inhibition and serum starvation and pulse-labelled for very short periods of time with 5-bromodeoxyuridine (5-BrdU). For the first time 5-BrdU has been employed to demonstrate the association of newly synthesized DNA with a nucleoskeleton. Immunofluorescence analysis using a monoclonal antibody to 5-BrdU revealed five different intranuclear staining patterns at different stages of the S phase. These patterns were observed also in intact cells and did not change during the matrix preparation steps which involve extraction with 2 M NaCl and DNase I digestion. Such an observation was also confirmed by spatial confocal microscopy studies. The intensity of fluorescence, which was evaluated by cytofluorometry, increased to reach a maximum during mid-S phase and then decreased. Because no significant difference was found in the time to label residual DNA of different 5-BrdU staining patterns, this strongly suggests that a different number of replicons is activated at different stages of the S phase. These results strengthen the hypothesis that eukaryotic DNA replication occurs in close association with an insoluble protein nuclear skeleton, which determines the three-dimensional spatial organization of chromosome duplication. |
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Authors:
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L M Neri; G Mazzotti; S Capitani; N M Maraldi; C Cinti; N Baldini; R Rana; A M Martelli |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Histochemistry Volume: 98 ISSN: 0301-5564 ISO Abbreviation: Histochemistry Publication Date: 1992 Aug |
Date Detail:
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Created Date: 1992-12-02 Completed Date: 1992-12-02 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0411300 Medline TA: Histochemistry Country: GERMANY |
Other Details:
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Languages: eng Pagination: 19-32 Citation Subset: IM |
Affiliation:
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Istituto di Anatomia Umana Normale, Universitá di Ferrara, Italy. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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3T3 Cells
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metabolism,
physiology Animals Bromodeoxyuridine* Cell Nucleus / physiology Culture Media, Serum-Free DNA / biosynthesis, genetics, metabolism DNA Replication / physiology* Mice Nuclear Matrix / metabolism, physiology* Replicon / physiology* |
| Chemical | |
Reg. No./Substance:
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0/Culture Media, Serum-Free; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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