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PubMed · 16777599

Sequence-resolved detection of pausing by single RNA polymerase molecules.

Kristina M Herbert·Arthur La Porta·Becky J Wong·Rachel A Mooney·Keir C Neuman·Robert Landick·Steven M Block
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Abstract

Transcriptional pausing by RNA polymerase (RNAP) plays an important role in the regulation of gene expression. Defined, sequence-specific pause sites have been identified biochemically. Single-molecule studies have also shown that bacterial RNAP pauses frequently during transcriptional elongation, but the relationship of these "ubiquitous" pauses to the underlying DNA sequence has been uncertain. We employed an ultrastable optical-trapping assay to follow the motion of individual molecules of RNAP transcribing templates engineered with repeated sequences carrying imbedded, sequence-specific pause sites of known regulatory function. Both the known and ubiquitous pauses appeared at reproducible locations, identified with base-pair accuracy. Ubiquitous pauses were associated with DNA sequences that show similarities to regulatory pause sequences. Data obtained for the lifetimes and efficiencies of pauses support a model where the transition to pausing branches off of the normal elongation pathway and is mediated by a common elemental state, which corresponds to the ubiquitous pause.

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Base SequenceDNA-Directed RNA PolymerasesPromoter Regions, GeneticSequence Analysis, DNATemplates, GeneticTranscription, GeneticJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov't

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Kristina M Herbert, Arthur La Porta, Becky J Wong, Rachel A Mooney, Keir C Neuman, Robert Landick, Steven M Block. 2006-06-16. Sequence-resolved detection of pausing by single RNA polymerase molecules.. https://doi.org/10.1016/j.cell.2006.04.032

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