Journal
JOURNAL OF CELL SCIENCE
Volume 117, Issue 19, Pages 4603-4614Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.01330
Keywords
large-scale chromatin organization; nuclear architecture; gene expression; mouse erythroleukemia cells; beta-globin gene
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Funding
- NIGMS NIH HHS [R01 GM058460, GM058460] Funding Source: Medline
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Previous work has demonstrated a more decondensed large-scale chromatin structure and a more internal nuclear position for gene-rich versus gene-poor chromosome regions. Here, we show that large-scale chromatin opening and changes in intranuclear positioning of chromosome regions can be induced by normal levels of endogenous transcription factors acting on mammalian regulatory sequences. We transfected mouse erythroleukemia cells with a 15 kbp plasmid containing a lac operator repeat plus beta-globin regulatory sequences driving a beta-galactosidase reporter gene. After green-fluorescent-protein/lac-repressor fusion-protein binding or after fluorescence in situ hybridization, the volume and location of the transgene array signal were measured. With both detection methods, we found that the volume was severalfold larger when transcription was on. While silent transgene arrays were located close to the nuclear membrane, we observed a significantly more internal position for the transcriptionally active state. Our results indicate that both large-scale chromatin decondensation and changes in nuclear positioning as observed for large, complex gene-rich chromosome regions can be reproduced by endogenous regulatory sequences acting within simple repetitive transgene arrays.
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