In vivo dynamics of RNA polymerase II transcription View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2007-09

AUTHORS

Xavier Darzacq, Yaron Shav-Tal, Valeria de Turris, Yehuda Brody, Shailesh M Shenoy, Robert D Phair, Robert H Singer

ABSTRACT

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. More... »

PAGES

796-806

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nsmb1280

DOI

http://dx.doi.org/10.1038/nsmb1280

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1051707754

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/17676063


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