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In vivo single-molecule dynamics of transcription of the viral T7 Phi 10 promoter in Escherichia coli

Research output: Chapter in Book/Report/Conference proceedingConference contributionScientificpeer-review

Details

Original languageEnglish
Title of host publicationThe 8th International Conference on Bioinformatics, Biocomputational Systems and Biotechnologies (BIOTECHNO2016)
PublisherIARIA
Pages9-15
ISBN (Electronic)978-1-61208-488-6
Publication statusPublished - 26 Jun 2016
Publication typeA4 Article in a conference publication
EventInternational Conference on Bioinformatics, Biocomputational Systems and Biotechnologies -
Duration: 1 Jan 2000 → …

Conference

ConferenceInternational Conference on Bioinformatics, Biocomputational Systems and Biotechnologies
Period1/01/00 → …

Abstract

We study the dynamics of transcription initiation of the T7 Phi 10 promoter as a function of temperature, using quantitative polymerase chain reaction (qPCR) and in vivo single-cell, single-ribonucleic acid (RNA) time-lapse microscopy. First, from the mean and squared coefficient of variation of the empirical distribution of intervals between consecutive RNA appearances in individual cells, we find that both the mean rate and noise in RNA production increase with temperature (from 20 °C to 43 °C). Next, the process is shown to be sub-Poissonian in all conditions, suggesting the existence of more than one rate-limiting step and absence of a significant ON-OFF mechanism. Next, from the kinetics of RNA production for varying amounts of T7 RNA polymerases, we find that as temperature increases, the fraction of time that the T7 RNA polymerase spends in open complex formation increases relative to the time to commit to closed complex formation, due to changes in the kinetics of open complex, closed complex, and reversibility of the closed complex formation. We conclude that the initiation kinetics of the T7 Phi 10 promoter changes with temperature due to changes in the kinetics of its rate-limiting steps.

Publication forum classification

Field of science, Statistics Finland