Document Detail

Nuclear matrix-bound replicational sites detected in situ by 5-bromodeoxyuridine.
MedLine Citation:
PMID:  1429014     Owner:  NLM     Status:  MEDLINE    
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.
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:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Histochemistry     Volume:  98     ISSN:  0301-5564     ISO Abbreviation:  Histochemistry     Publication Date:  1992 Aug 
Date Detail:
Created Date:  1992-12-02     Completed Date:  1992-12-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0411300     Medline TA:  Histochemistry     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  19-32     Citation Subset:  IM    
Istituto di Anatomia Umana Normale, Universitá di Ferrara, Italy.
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MeSH Terms
3T3 Cells / metabolism,  physiology
Cell Nucleus / physiology
Culture Media, Serum-Free
DNA / biosynthesis,  genetics,  metabolism
DNA Replication / physiology*
Nuclear Matrix / metabolism,  physiology*
Replicon / physiology*
Reg. No./Substance:
0/Culture Media, Serum-Free; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA

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