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PMID: 2910875 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mapping replicational sites in the eucaryotic cell nucleus.

The Journal of cell biology ·Vol. 108 ·No. 1 ·1989-01-00 ·Pages 1-11

Nakayasu H, Berezney R

Abstract

We have used fluorescent microscopy to map DNA replication sites in the interphase cell nucleus after incorporation of biotinylated dUTP into permeabilized PtK-1 kangaroo kidney or 3T3 mouse fibroblast cells. Discrete replication granules were found distributed throughout the nuclear interior and along the periphery. Three distinct patterns of replication sites in relationship to chromatin domains in the cell nucleus and the period of S phase were detected and termed type I (early to mid S), type II (mid to late S) and type III (late S). Similar patterns were seen with in vivo replicated DNA using antibodies to 5-bromodeoxyuridine. Extraction of the permeabilized cells with DNase I and 0.2 M ammonium sulfate revealed a striking maintenance of these replication granules and their distinct intranuclear arrangements with the remaining nuclear matrix structures despite the removal of greater than 90% of the total nuclear DNA. The in situ prepared nuclear matrix structures also incorporated biotinylated dUTP into replication granules that were indistinguishable from those detected within the intact nucleus.

MeSH Terms
Animals Biotin/analogs & derivatives,metabolism Cell Line Cell Nucleus/metabolism,ultrastructure Chromatin/metabolism,ultrastructure DNA/biosynthesis DNA Replication Deoxyuracil Nucleotides/metabolism Interphase Microscopy, Fluorescence
Chemicals
Chromatin Deoxyuracil Nucleotides Biotin biotin-11-dUTP DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nakayasu H
Department of Biological Sciences, State University of New York, Buffalo 14260.
Berezney R
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.108.1.1
Published
1989-01-00
Pages
1-11
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115357
Subset
IM
Grants
NIGMS NIH HHS · GM-23922 · United States
External Links
PubMed source DOI: 10.1083/jcb.108.1.1 PMC Full Text View journal details
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