A strategy for dissecting the kinetics of transcription repression mechanisms

Cristina S. D. Palma, Sofia Startceva, Ramakanth Neeli-Venkata, Marzieh Zare, Nadia S. M. Gonçalves, Jose M. Fonseca, Samuel M. D. Oliveira, Andre S. Ribeiro

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

1 Citation (Scopus)

Abstract

Promoters in Escherichia coli include an ‘OFF’ state, during which transcription is halted. Here, we propose a novel empirical method for assessing the time-length spent by promoters in this state. It relies on direct measurements of RNA production kinetics at the single molecule level at different induction levels, followed by an estimation of the RNA production rate under infinite induction, which is then compared to this rate under real, maximum induction. We apply it to the LacO3O1 promoter and infer that, under full induction, on average, 15% of the time between successful transcription events is spent in the OFF state. We verify this result by comparing the kinetics of a mutant strain lacking repressor molecules with that of the inferred rate under infinite induction. We expect this strategy of dissecting the kinetics of transcription repression to be applicable to a wide number of promoters in E. coli.

Original languageEnglish
Title of host publicationEMBEC and NBC 2017 - Joint Conference of the European Medical and Biological Engineering Conference EMBEC 2017 and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics, NBC 2017
PublisherSpringer Verlag
Pages1097-1100
Number of pages4
Volume65
ISBN (Print)9789811051210
DOIs
Publication statusPublished - 1 Jan 2017
EventJoint Conference of the European Medical and Biological Engineering Conference, EMBEC 2017 and Nordic-Baltic Conference on Biomedical Engineering and Medical Physics, NBC 2107 - Tampere, Finland
Duration: 11 Jun 201715 Jun 2017

Conference

ConferenceJoint Conference of the European Medical and Biological Engineering Conference, EMBEC 2017 and Nordic-Baltic Conference on Biomedical Engineering and Medical Physics, NBC 2107
CountryFinland
CityTampere
Period11/06/1715/06/17

Keywords

  • Induction
  • OFF state
  • Transcription
  • τ plot

Fingerprint Dive into the research topics of 'A strategy for dissecting the kinetics of transcription repression mechanisms'. Together they form a unique fingerprint.

Cite this