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Protection of Escherichia coli cells against extreme turgor by activation of MscS and MscL mechanosensitive channels: identification of genes required for MscS activity.

TLDR
The characterization of a new gene family required for MscS function, YggB and KefA, is reported, which has enabled a rigorous test of the role of the channels.
Abstract
Mechanosensitive channels are ubiquitous amongst bacterial cells and have been proposed to have major roles in the adaptation to osmotic stress, in particular in the management of transitions from high to low osmolarity environments. Electrophysiological measurements have identified multiple channels in Escherichia coli cells. One gene, mscL, encoding a large conductance channel has previously been described, but null mutants were without well-defined phenotypes. Here, we report the characterization of a new gene family required for MscS function, YggB and KefA, which has enabled a rigorous test of the role of the channels. The channel determined by KefA does not appear to have a major role in managing the transition from high to low osmolarity. In contrast, analysis of mutants of E.coli lacking YggB and MscL shows that mechanosensitive channels are designed to open at a pressure change just below that which would cause cell disruption leading to death.

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Osmotic Stress Signaling and Osmoadaptation in Yeasts

TL;DR: An integrated understanding of osmoadaptation requires not only knowledge of the function of many uncharacterized genes but also further insight into the time line of events, their interdependence, their dynamics, and their spatial organization as well as the importance of subtle effects.
Journal ArticleDOI

Structure, Function, and Evolution of Bacterial ATP-Binding Cassette Systems

TL;DR: The availability of an increasing number of high-resolution structures has provided a valuable framework for interpretation of recent studies, and realistic models have been proposed to explain how these fascinating molecular machines use complex dynamic processes to fulfill their numerous biological functions.
Journal ArticleDOI

Molecular basis of mechanotransduction in living cells

TL;DR: The simplest cell-like structure, the lipid bilayer vesicle, can respond to mechanical deformation by elastic membrane dilation/thinning and curvature changes and changes in local membrane curvature may shift the equilibrium between channel conformations.
Journal ArticleDOI

Bacterial osmoadaptation: the role of osmolytes in bacterial stress and virulence

TL;DR: The molecular mechanisms governing the accumulation of these compounds, both in Gram-positive and Gram-negative bacteria, are reviewed, focusing specifically on the regulation of their transport/synthesis systems and the ability of these systems to sense and respond to changes in the osmolarity of the extracellular environment.
References
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Journal ArticleDOI

Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.

TL;DR: A new criterion for triggering the extension of word hits, combined with a new heuristic for generating gapped alignments, yields a gapped BLAST program that runs at approximately three times the speed of the original.
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A simple method for displaying the hydropathic character of a protein

TL;DR: A computer program that progressively evaluates the hydrophilicity and hydrophobicity of a protein along its amino acid sequence has been devised and its simplicity and its graphic nature make it a very useful tool for the evaluation of protein structures.
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Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.

TL;DR: The extracellular patch clamp method, which first allowed the detection of single channel currents in biological membranes, has been further refined to enable higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches.
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Site-directed mutagenesis by overlap extension using the polymerase chain reaction.

TL;DR: In this paper, complementary oligodeoxyribonucleotide (oligo) primers and the polymerase chain reaction are used to generate two DNA fragments having overlapping ends, and these fragments are combined in a subsequent 'fusion' reaction in which the overlapping ends anneal, allowing the 3' overlap of each strand to serve as a primer for the three' extension of the complementary strand.
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A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria

TL;DR: In this paper, a new vector strategy for the insertion of foreign genes into the genomes of gram negative bacteria not closely related to Escherichia coli was developed, which can utilize any gram negative bacterium as a recipient for conjugative DNA transfer.
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