Journal
CELL
Volume 116, Issue 5, Pages 683-698Publisher
CELL PRESS
DOI: 10.1016/S0092-8674(04)00171-0
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Funding
- NIBIB NIH HHS [R01 EB002060] Funding Source: Medline
- NIGMS NIH HHS [GM067728, GM42694, R01 GM067728, R01 GM042694] Funding Source: Medline
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We have developed an inducible system to visualize gene expression at the levels of DNA, RNA and protein in living cells. The system is composed of a 200 copy transgene array integrated into a euchromatic region of chromosome 1 in human U20S cells. The condensed array is heterochromatic as it is associated with HP1, histone H3 methylated at lysine 9, and several histone methyltransferases. Upon transcriptional induction, HP1alpha is depleted from the locus and the histone variant H3.3 is deposited suggesting that histone exchange is a mechanism through which heterochromatin is transformed into a transcriptionally active state. RNA levels at the transcription site increase immediately after the induction of transcription and the rate of synthesis slows over time. Using this system, we are able to correlate changes in chromatin structure with the progression of transcriptional activation allowing us to obtain a real-time integrative view of gene expression.
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