TY - JOUR
T1 - From silencing to gene expression
T2 - Real-time analysis in single cells
AU - Janicki, Susan M.
AU - Tsukamoto, Toshiro
AU - Salghetti, Simone E.
AU - Tansey, William P.
AU - Sachidanandam, Ravi
AU - Prasanth, Kannanganattu V.
AU - Ried, Thomas
AU - Shav-Tal, Yaron
AU - Bertrand, Edouard
AU - Singer, Robert H.
AU - Spector, David L.
N1 - Funding Information:
We would like to thank Carolyn Dong for designing the plasmid schematic in Figure 1 ; Supriya Prasanth for help with the Northern blot analysis; Steve Henikoff for insightful discussions about histone exchange; David Allis, Thomas Jenuwein, Frank Rauscher, Yoshihiro Nakatani, and James Manley for their generous gifts of antibodies. We would also like to thank Edith Heard and members of the Spector laboratory for critical review of the manuscript. Supported by grant 42694 from NIGMS/NIH to D.L.S. and 067728 to W.P.T. W.P.T. is a Leukemia and Lymphoma Society Scholar.
PY - 2004/3/5
Y1 - 2004/3/5
N2 - 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 U2OS 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, HP1α 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.
AB - 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 U2OS 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, HP1α 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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U2 - 10.1016/S0092-8674(04)00171-0
DO - 10.1016/S0092-8674(04)00171-0
M3 - Article
C2 - 15006351
AN - SCOPUS:12144289862
SN - 0092-8674
VL - 116
SP - 683
EP - 698
JO - Cell
JF - Cell
IS - 5
ER -