Source:http://linkedlifedata.com/resource/pubmed/id/17676063
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
9
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pubmed:dateCreated |
2007-9-7
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pubmed:abstractText |
We imaged transcription in living cells using a locus-specific reporter system, which allowed precise, single-cell kinetic measurements of promoter binding, initiation and elongation. Photobleaching of fluorescent RNA polymerase II revealed several kinetically distinct populations of the enzyme interacting with a specific gene. Photobleaching and photoactivation of fluorescent MS2 proteins used to label nascent messenger RNAs provided sensitive elongation measurements. A mechanistic kinetic model that fits our data was validated using specific inhibitors. Polymerases elongated at 4.3 kilobases min(-1), much faster than previously documented, and entered a paused state for unexpectedly long times. Transcription onset was inefficient, with only 1% of polymerase-gene interactions leading to completion of an mRNA. Our systems approach, quantifying both polymerase and mRNA kinetics on a defined DNA template in vivo with high temporal resolution, opens new avenues for studying regulation of transcriptional processes in vivo.
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pubmed:grant | |
pubmed:commentsCorrections | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
1545-9993
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
14
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
796-806
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pubmed:meshHeading |
pubmed-meshheading:17676063-Base Sequence,
pubmed-meshheading:17676063-Cell Line, Tumor,
pubmed-meshheading:17676063-DNA Primers,
pubmed-meshheading:17676063-Humans,
pubmed-meshheading:17676063-In Situ Hybridization, Fluorescence,
pubmed-meshheading:17676063-Kinetics,
pubmed-meshheading:17676063-Phosphorylation,
pubmed-meshheading:17676063-Photochemistry,
pubmed-meshheading:17676063-RNA Polymerase II,
pubmed-meshheading:17676063-Transcription, Genetic
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pubmed:year |
2007
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pubmed:articleTitle |
In vivo dynamics of RNA polymerase II transcription.
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pubmed:affiliation |
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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