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PMID: 1740468 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences.

The Journal of cell biology ·Vol. 116 ·No. 5 ·1992-03-00 ·Pages 1095-110

O'Keefe RT, Henderson SC, Spector DL

Abstract

Five distinct patterns of DNA replication have been identified during S-phase in asynchronous and synchronous cultures of mammalian cells by conventional fluorescence microscopy, confocal laser scanning microscopy, and immunoelectron microscopy. During early S-phase, replicating DNA (as identified by 5-bromodeoxyuridine incorporation) appears to be distributed at sites throughout the nucleoplasm, excluding the nucleolus. In CHO cells, this pattern of replication peaks at 30 min into S-phase and is consistent with the localization of euchromatin. As S-phase continues, replication of euchromatin decreases and the peripheral regions of heterochromatin begin to replicate. This pattern of replication peaks at 2 h into S-phase. At 5 h, perinucleolar chromatin as well as peripheral areas of heterochromatin peak in replication. 7 h into S-phase interconnecting patches of electron-dense chromatin replicate. At the end of S-phase (9 h), replication occurs at a few large regions of electron-dense chromatin. Similar or identical patterns have been identified in a variety of mammalian cell types. The replication of specific chromosomal regions within the context of the BrdU-labeling patterns has been examined on an hourly basis in synchronized HeLa cells. Double labeling of DNA replication sites and chromosome-specific alpha-satellite DNA sequences indicates that the alpha-satellite DNA replicates during mid S-phase (characterized by the third pattern of replication) in a variety of human cell types. Our data demonstrates that specific DNA sequences replicate at spatially and temporally defined points during the cell cycle and supports a spatially dynamic model of DNA replication.

MeSH Terms
Animals Bromodeoxyuridine CHO Cells Cell Compartmentation Cell Nucleus/metabolism Centromere/metabolism Cricetinae DNA Replication DNA, Satellite/analysis Female HeLa Cells Humans Male S Phase Tumor Cells, Cultured
Chemicals
DNA, Satellite Bromodeoxyuridine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
O'Keefe R T
Cold Spring Harbor Laboratory, New York 11724-2217.
Henderson S C
Spector D L
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.116.5.1095
Published
1992-03-00
Pages
1095-110
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289349
Subset
IM
Grants
NIGMS NIH HHS · R01 GM042694 · United States
NCI NIH HHS · 5P30 CA45508-03 · United States
NIGMS NIH HHS · GM42694 · United States
External Links
PubMed source DOI: 10.1083/jcb.116.5.1095 PMC Full Text View journal details
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