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Title: Single cell super-resolution imaging of E. coli OmpR during environmental stress
Authors: Foo, Y.H 
Spahn, C
Zhang, H 
Heilemann, M
Kenney, L.J 
Keywords: amino acid
hybrid protein
outer membrane protein regulator
protein envz
protein histidine kinase
RNA polymerase
unclassified drug
bacterial DNA
bacterial protein
envZ protein, E coli
Escherichia coli protein
hybrid protein
multienzyme complex
osmolarity response regulator proteins
outer membrane protein
protein binding
transactivator protein
bacterial cell
bacterial chromosome
binding affinity
controlled study
environmental stress
Escherichia coli
inner membrane
priority journal
protein binding
protein expression
protein localization
quantitative analysis
Western blotting
osmotic pressure
physiological stress
protein quaternary structure
signal transduction
single cell analysis
Bacterial Outer Membrane Proteins
Bacterial Proteins
Biomechanical Phenomena
Chromosomes, Bacterial
DNA, Bacterial
Escherichia coli
Escherichia coli Proteins
Hydrogen-Ion Concentration
Multienzyme Complexes
Osmotic Pressure
Protein Binding
Protein Structure, Quaternary
Recombinant Fusion Proteins
Signal Transduction
Single-Cell Analysis
Spectrometry, Fluorescence
Stress, Physiological
Issue Date: 2015
Citation: Foo, Y.H, Spahn, C, Zhang, H, Heilemann, M, Kenney, L.J (2015). Single cell super-resolution imaging of E. coli OmpR during environmental stress. Integrative Biology (United Kingdom) 7 (10) : 1297-1308. ScholarBank@NUS Repository.
Abstract: Two-component signaling systems are a major strategy employed by bacteria, and to some extent, yeast and plants, to respond to environmental stress. The EnvZ/OmpR system in E. coli responds to osmotic and acid stress and is responsible for regulating the protein composition of the outer membrane. EnvZ is a histidine kinase located in the inner membrane. Upon activation, it is autophosphorylated by ATP and subsequently, it activates OmpR. Phosphorylated OmpR binds with high affinity to the regulatory regions of the ompF and ompC porin genes to regulate their transcription. We set out to visualize these two-components in single bacterial cells during different environmental stress conditions and to examine the subsequent modifications to the bacterial nucleoid as a result. We created a chromosomally-encoded, active, fluorescent OmpR-PAmCherry fusion protein and compared its expression levels with RNA polymerase. Quantitative western blotting had indicated that these two proteins were expressed at similar levels. From our images, it is evident that OmpR is significantly less abundant compared to RNA polymerase. In cross-sectional axial images, we observed OmpR molecules closely juxtaposed near the inner membrane during acidic and hyposomotic growth. In acidic conditions, the chromosome was compacted. Surprisingly, under acidic conditions, we also observed evidence of a spatial correlation between the DNA and the inner membrane, suggesting a mechanical link through an active DNA-OmpR-EnvZ complex. This work represents the first direct visualization of a response regulator with respect to the bacterial chromosome. © 2015 The Royal Society of Chemistry.
Source Title: Integrative Biology (United Kingdom)
ISSN: 17579694
DOI: 10.1039/c5ib00077g
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