Dynamics and stoichiometry of a regulated enhancer-binding protein in live Escherichia coli cells

Parul Mehta, Goran Jovanovic, Tchern Lenn, Andreas Bruckbauer, Christoph Engl, Liming Ying () and Martin Buck ()
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Parul Mehta: Imperial College London
Goran Jovanovic: Imperial College London
Tchern Lenn: Imperial College London
Andreas Bruckbauer: Lymphocyte Interaction Laboratory, London Research Institute, Cancer Research UK
Christoph Engl: Imperial College London
Liming Ying: Faculty of Medicine, Molecular Medicine, National Heart and Lung Institute, Imperial College London
Martin Buck: Imperial College London

Nature Communications, 2013, vol. 4, issue 1, 1-9

Abstract: Abstract Bacterial enhancer-dependent transcription systems support major adaptive responses and offer a singular paradigm in gene control analogous to complex eukaryotic systems. Here we report new mechanistic insights into the control of one-membrane stress-responsive bacterial enhancer-dependent system. Using millisecond single-molecule fluorescence microscopy of live cells we determine the localizations, two-dimensional diffusion dynamics and stoichiometries of complexes of the bacterial enhancer-binding ATPase PspF during its action at promoters as regulated by inner membrane interacting negative controller PspA. We establish that a stable repressive PspF–PspA complex is located in the nucleoid, transiently communicating with the inner membrane via PspA. The PspF as a hexamer stably binds only one of the two psp promoters at a time, suggesting that psp promoters will fire asynchronously and cooperative interactions of PspF with the basal transcription complex influence dynamics of the PspF hexamer–DNA complex and regulation of the psp promoters.

Date: 2013
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DOI: 10.1038/ncomms2997

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