DNA replication and nuclear architecture

• Published in: Journal of cellular biochemistry Vol. 85; no. 1; pp. 1 - 9
• Main Authors: Jaunin, Françoise, Fakan, Stanislav
• Format: Journal Article
• Language: English
• Published: New York Wiley Subscription Services, Inc., A Wiley Company 2002
• Subjects: Animals; Cell Nucleus - genetics; Cell Nucleus - ultrastructure; Chromatin - genetics; Chromatin - ultrastructure; DNA Replication; electron microscopy; fluorescence microscopy; Humans; Microscopy, Electron - methods; Microscopy, Fluorescence - methods; nuclear domains; replication foci; replication sites; Replicon - genetics; Replicon - physiology; S Phase - physiology
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.10115

The model of in situ DNA replication provided by immunofluorescence and confocal imaging is compared with observations obtained by electron microscopic studies. Discrepancies between both types of observations call into question the replication focus as a persistent nuclear structure and as a replication entity where DNA replication takes place. Most electron microscopic analyses reveal that replication sites are confined to dispersed chromatin areas at the periphery of condensed chromatin, and the distribution of replication factors exhibits the same localization pattern. Moreover, rapid migration of newly synthesized DNA from the replication sites towards the interior of condensed chromatin regions obviously takes place during S‐phase. It implies modifications of replication domains, hardly detectable by fluorescence microscopy. The confrontation of different observations carried out at light microscopic or electron microscopic levels of resolution lead to a conclusion that a combination of in vivo fluorescence analysis with a subsequent ultrastructural investigation performed on the same cells will represent an optimal approach in future studies of nuclear functions in situ. J. Cell. Biochem. 85: 1–9, 2002. © 2002 Wiley‐Liss, Inc.