Journal of Cell Science, Vol 107, Issue 2 425-434, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Replication and transcription sites are colocalized in human cells

AB Hassan, RJ Errington, NS White, DA Jackson and PR Cook
CRC Nuclear Structure and Function Research Group, Sir William Dunn School of Pathology, University of Oxford, UK.

HeLa cells synchronized at different stages of the cell cycle were permeabilized and incubated with analogues of nucleotide triphosphates; then sites of incorporation were immunolabeled with the appropriate fluorescent probes. Confocal microscopy showed that sites of replication and transcription were not diffusely spread throughout nuclei, reflecting the distribution of euchromatin; rather, they were concentrated in 'foci' where many polymerases act together. Transcription foci aggregated as cells progressed towards the G1/S boundary; later they dispersed and became more diffuse. Replication was initiated only at transcription sites; later, when heterochromatin was replicated in enlarged foci, these remained sites of transcription. This illustrates the dynamic nature of nuclear architecture and suggests that transcription may be required for the initiation of DNA synthesis.
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