Organization of early and late replicating DNA in human chromosome territories.
It has been suggested that DNA organized into replication foci during S-phase remains stably aggregated in non-S-phase cells and that these stable aggregates provide fundamental units of nuclear or chromosome architecture [C. Meng and R. Berezney (1991) J. Cell Biol. 115, 95a; E. Sparvoli et al. (1994) J. Cell Sci. 107, 3097-3103; D. A. Jackson and A. Pombo (1998) J. Cell Biol. 140, 1285-1295; D. Zink et al. (1998) Hum. Genet. 112, 241-251]. To test this hypothesis, early and late replicating DNA of human diploid fibroblasts was labeled specifically by incorporating two different thymidine analogs [J. Aten (1992) Histochem. J. 24, 251-259; A. E. Visser (1998) Exp. Cell Res. 243, 398-407], during distinct time segments of S-phase. On mitotic chromosomes the amount and spatial distribution of early and late replicating DNA corresponded to R/G-banding patterns. After labeling cells were grown for several cell cycles. During this growth period individual replication labeled chromosomes were distributed into an environment of unlabeled chromosomes. The nuclear territories of chromosomes 13 and 15 were identified by additional chromosome painting. The distribution of early and late replicating DNA was analyzed for both chromosomes in quiescent (G0) cells or at G1. Early and late replicating DNA occupied distinct foci within chromosome territories, displaying a median overlap of only 5-10%. There was no difference in this regard between G1 and G0 cells. Chromosome 13 and 15 territories displayed a similar structural rearrangement in G1 cells compared to G0 cells resulting in the compaction of the territories. The findings demonstrate that early and late replicating foci are maintained during subsequent cell cycles as distinctly separated units of chromosome organization. These findings are compatible with the hypothesis that DNA organized into replicon clusters remains stably aggregated in non-S-phase cells.
Zink
D
D
Institut für Anthropologie und Humangenetik, LMU München, Goethestrasse 31, München, D-80336, Germany. dani.zink@lrz.uni-muenchen.de
Bornfleth
H
H
Visser
A
A
Cremer
C
C
Cremer
T
T
D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000378:metabolism*; D002882:Chromosomes, Human, Pair 13 / Q000378:metabolism; D002884:Chromosomes, Human, Pair 15 / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication / Q000235:genetics* / Q000502:physiology*; D003857:Deoxyuridine / Q000031:analogs & derivatives; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007065:Idoxuridine; D007399:Interphase / Q000235:genetics / Q000502:physiology; D016192:Resting Phase, Cell Cycle / Q000235:genetics / Q000502:physiology; D013194:Staining and Labeling
C012244:5-chloro-2'-deoxyuridine; D004247:DNA; D007065:Idoxuridine; D003857:Deoxyuridine