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- W2003253968 abstract "How efficient is transcription in vivo? Recent fluorescence microscopy studies have begun to confront this question by measuring the binding times of RNA polymerase II (pol II) and other components of the transcription machinery in single living cells. This has revealed an extremely dynamic transcription machine that appears to operate with components in continual flux. However, it remains debatable whether or not these rapid dynamics lead to efficient transcription, mainly because it has been difficult to distinguish different phases of the transcription cycle in vivo. For example, using GFP it is possible to visualize the movement of pol II, but directly distinguishing the uninitiated fraction from the initiated or elongating fractions has not yet been possible. To overcome this difficulty, we have loaded fluorescent antibody fragments (Fab) against unphosphorylated and phosphorylated forms of pol II into living cells containing an inducible tandem gene array. Using this unique system, we are quantifying the accumulation of uninitiated (unphosphorylated), initiated (ser 5 phosphorylated), and elongating (ser 2 phosphorylated) forms of pol II to the gene array after induction. Our observations suggest transcription is quite efficient, with pol II being recruited to activated genes within ∼3 minutes time, after which ∼70% are initiated within 30 seconds and ∼35% proceed to elongation within another 30 seconds. We are currently investigating how these transcription dynamics correlate with histone modification dynamics at the gene array." @default.
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- W2003253968 date "2012-01-01" @default.
- W2003253968 modified "2023-09-28" @default.
- W2003253968 title "Visualizing the Transcription Cycle of Endogenous RNA Polymerase II in Single Living Cells" @default.
- W2003253968 doi "https://doi.org/10.1016/j.bpj.2011.11.1252" @default.
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