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© The Rockefeller University Press, 0021-9525/1997//1597 $5.00
The Journal of Cell Biology, Volume 139, Number 7, , 1997 1597-1610

Article

Spatial Organization of Large-Scale Chromatin Domains in the Nucleus: A Magnified View of Single Chromosome Territories



João Ferreira*, Giovanni Paolella{ddagger}, Carlos Ramos*, and Angus I. Lamond§

* Institute of Histology, Faculty of Medicine, 1699 Lisboa codex, Portugal; {ddagger} Dipartimento di Biochimica e Mediche, via Sergio Pansini 5, 80131 Napoli, Italy; and § Department of Biochemistry, University of Dundee, Dundee DD14HN, Scotland, UK.

We have analyzed the spatial organization of large scale chromatin domains in chinese hamster fibroblast, human lymphoid (IM-9), and marsupial kidney epithelial (PtK) cells by labeling DNA at defined stages of S phase via pulsed incorporation of halogenated deoxynucleosides. Most, if not all, chromosomes contribute multiple chromatin domains to both peripheral and internal nucleoplasmic compartments. The peripheral compartment contains predominantly late replicating G/Q bands, whereas early replicating R bands preferentially localize to the internal nucleoplasmic compartment. During mitosis, the labeled chromatin domains that were separated in interphase form a pattern of intercalated bands along the length of each metaphase chromosome. The transition from a banded (mitotic) to a compartmentalized (interphasic) organization of chromatin domains occurs during the late telophase/early G1 stage and is independent of transcriptional activation of the genome. Interestingly, generation of micronuclei with a few chromosomes showed that the spatial separation of early and late replicating chromatin compartments is recapitulated independently of chromosome number, even in micronuclei containing only a single chromosome. Our data strongly support the notion that the compartmentalization of large-scale (band size) chromatin domains seen in the intact nucleus is a magnified image of a similar compartmentalization occurring in individual chromosome territories.

Abbreviations used in this paper: 3D; three-dimensional CHF, chinese hamster fibroblast; PtK, marsupial kidney epithelial; SAR, scaffold associated region.

J. Ferreira was supported by a Calouste Gulbenkian Foundation fellowship.

Address all correspondence to Dr. Angus I. Lamond, Department of Biochemistry, University of Dundee, Dundee DD14HN, Scotland, UK. Tel: 1-382-345-473. Fax: 1-382-345-695. E-mail: a.i.lamond{at}dundee.ac.uk


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