Spatial distributions of early and late replicating chromatin in interphase chromosome territories.
The surface area of chromosome territories has been suggested as a preferred site for genes, specific RNAs, and accumulations of splicing factors. Here, we investigated the localization of sites of replication within individual chromosome territories. In vivo replication labeling with thymidine analogues IdUrd and CldUrd was combined with chromosome painting by fluorescent in situ hybridization on three-dimensionally preserved human fibroblast nuclei. Spatial distributions of replication labels over the chromosome territory, as well as the territory volume and shape, were determined by 3D image analysis. During late S-phase a previously observed shape difference between the active and inactive X-chromosome in female cells was maintained, while the volumes of the two territories did not differ significantly. Domains containing early or mid to late replicating chromatin were distributed throughout territories of chromome 8 and the active X. In the inactive X-chromosome early replicating chromatin was observed preferentially near the territory surface. Most important, we established that the process of replication takes place in foci throughout the entire chromosome territory volume, in early as well as in late S-phase. This demonstrates that activity of macromolecular enzyme complexes takes place throughout chromosome territories and is not confined to the territory surface as suggested previously.
Visser
A E
AE
Department of Cell Biology, Academic Medical Center, University of Amsterdam, Amsterdam, 1105 AZ, The Netherlands. a.e.visser@amc.uva.nl
Eils
R
R
Jauch
A
A
Little
G
G
Bakker
P J
PJ
Cremer
T
T
Aten
J A
JA
D002478:Cells, Cultured; D002843:Chromatin / Q000502:physiology*; D020223:Chromosome Painting; D002875:Chromosomes; D002877:Chromosomes, Human / Q000502:physiology*; D004261:DNA Replication; D005260:Female; D005347:Fibroblasts; D006801:Humans; D007399:Interphase; D008938:Mitosis; D016196:S Phase; D014960:X Chromosome
D002843:Chromatin