Tampere University of Technology

TUTCRIS Research Portal

Dissecting the stochastic transcription initiation process in live Escherichia coli

Research output: Contribution to journalArticleScientificpeer-review

Details

Original languageEnglish
Pages (from-to)203-214
JournalDNA Research
Volume23
Issue number3
DOIs
Publication statusPublished - 29 Mar 2016
Publication typeA1 Journal article-refereed

Abstract

We investigate the hypothesis that, in Escherichia coli, while the concentration of RNA polymerases differs in different growth conditions, the fraction of RNA polymerases free for transcription remains approximately constant within a certain range of these conditions. After establishing this, we apply a standard model-fitting procedure to fully characterize the in vivo kinetics of the rate-limiting steps in transcription initiation of the Plac/ara-1 promoter from distributions of intervals between transcription events in cells with different RNA polymerase concentrations. We find that, under full induction, the closed complex lasts ∼788 s while subsequent steps last ∼193 s, on average. We then establish that the closed complex formation usually occurs multiple times prior to each successful initiation event. Furthermore, the promoter intermittently switches to an inactive state that, on average, lasts ∼87 s. This is shown to arise from the intermittent repression of the promoter by LacI. The methods employed here should be of use to resolve the rate-limiting steps governing the in vivo dynamics of initiation of prokaryotic promoters, similar to established steady-state assays to resolve the in vitro dynamics.

Publication forum classification

Field of science, Statistics Finland