Kinetics of the cellular intake of a gene expression inducer at high concentrations

Mol Biosyst. 2015 Sep;11(9):2579-87. doi: 10.1039/c5mb00244c. Epub 2015 Jul 30.

Abstract

From in vivo single-event measurements of the transient and steady-state transcription activity of a single-copy lac-ara-1 promoter in Escherichia coli, we characterize the intake kinetics of its inducer (IPTG) from the media. We show that the empirical data are well-fit by a model of intake assuming a bilayer membrane, with the passage through the second layer being rate-limiting, coupled to a stochastic, sub-Poissonian, multi-step transcription process. Using this model, we show that for a wide range of extracellular inducer levels (up to 1.25 mM) the intake process is diffusive-like, suggesting unsaturated membrane permeability. Inducer molecules travel from the periplasm to the cytoplasm in, on average, 31.7 minutes, strongly affecting cells' response time. The novel methodology followed here should aid the study of cellular intake mechanisms at the single-event level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Biological Transport
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Gene Expression Regulation, Bacterial / drug effects*
  • Intracellular Membranes / metabolism
  • Isopropyl Thiogalactoside / metabolism
  • Isopropyl Thiogalactoside / pharmacology
  • Models, Biological
  • Operon
  • Repressor Proteins / metabolism
  • Transcription, Genetic

Substances

  • Repressor Proteins
  • Isopropyl Thiogalactoside