YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity

Ulrike Schumann, Michelle D. Edwards, Tim Rasmussen, Wendy Bartlett, Pieter van West, Ian Rylance Booth

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

We describe a mechanosensitive (MS) channel that has mechanosensitive channel of miniconductance (MscM) activity, and displays unique properties with respect to gating. Mechanosensitive channels respond to membrane tension, are ubiquitous from bacteria to man, and exhibit a great diversity in structure and function. These channels protect Bacteria and Archaea against hypoosmotic shock and are critical determinants of shape in chloroplasts. Given the dominant roles played in bacteria by the mechanosensitive channel of small conductance (MscS) and the mechanosensitive channel of large conductance (MscL), the role of the multiple MS channel homologs observed in most organisms remains obscure. Here we demonstrate that a MscS homolog, YbdG, extends the range of hypoosmotic shock that Escherichia coli cells can survive, but its expression level is insufficient to protect against severe shocks. Overexpression of the YbdG protein provides complete protection. Transcription and translation of the ybdG gene are enhanced by osmotic stress consistent with a role for the protein in survival of hypoosmotic shock. Measurement of the conductance of the native channel by standard patch clamp methods was not possible. However, a fully functional YbdG mutant channel, V229A, exhibits a conductance in membrane patches consistent with MscM activity. We find that MscM activities arise from more than one gene product because ybdG deletion mutants still exhibit an occasional MscMlike conductance. We propose that ybdG encodes a low- abundance MscM- type MS channel, which in cells relieves low levels of membrane tension, obviating the need to activate the major MS channels, MscS and MscL.

Original languageEnglish
Pages (from-to)12664-12669
Number of pages6
JournalPNAS
Volume107
Issue number28
DOIs
Publication statusPublished - 13 Jul 2010

Keywords

  • MscS
  • osmoregulation
  • activated channels
  • stationary phase
  • ion channels
  • membrane
  • proteins
  • gene
  • identification
  • expression
  • pressure
  • RPOS

Cite this

Schumann, U., Edwards, M. D., Rasmussen, T., Bartlett, W., van West, P., & Booth, I. R. (2010). YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity. PNAS, 107(28), 12664-12669. https://doi.org/10.1073/pnas.1001405107

YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity. / Schumann, Ulrike; Edwards, Michelle D.; Rasmussen, Tim; Bartlett, Wendy; van West, Pieter; Booth, Ian Rylance.

In: PNAS, Vol. 107, No. 28, 13.07.2010, p. 12664-12669.

Research output: Contribution to journalArticle

Schumann, U, Edwards, MD, Rasmussen, T, Bartlett, W, van West, P & Booth, IR 2010, 'YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity', PNAS, vol. 107, no. 28, pp. 12664-12669. https://doi.org/10.1073/pnas.1001405107
Schumann, Ulrike ; Edwards, Michelle D. ; Rasmussen, Tim ; Bartlett, Wendy ; van West, Pieter ; Booth, Ian Rylance. / YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity. In: PNAS. 2010 ; Vol. 107, No. 28. pp. 12664-12669.
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AU - Bartlett, Wendy

AU - van West, Pieter

AU - Booth, Ian Rylance

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N2 - We describe a mechanosensitive (MS) channel that has mechanosensitive channel of miniconductance (MscM) activity, and displays unique properties with respect to gating. Mechanosensitive channels respond to membrane tension, are ubiquitous from bacteria to man, and exhibit a great diversity in structure and function. These channels protect Bacteria and Archaea against hypoosmotic shock and are critical determinants of shape in chloroplasts. Given the dominant roles played in bacteria by the mechanosensitive channel of small conductance (MscS) and the mechanosensitive channel of large conductance (MscL), the role of the multiple MS channel homologs observed in most organisms remains obscure. Here we demonstrate that a MscS homolog, YbdG, extends the range of hypoosmotic shock that Escherichia coli cells can survive, but its expression level is insufficient to protect against severe shocks. Overexpression of the YbdG protein provides complete protection. Transcription and translation of the ybdG gene are enhanced by osmotic stress consistent with a role for the protein in survival of hypoosmotic shock. Measurement of the conductance of the native channel by standard patch clamp methods was not possible. However, a fully functional YbdG mutant channel, V229A, exhibits a conductance in membrane patches consistent with MscM activity. We find that MscM activities arise from more than one gene product because ybdG deletion mutants still exhibit an occasional MscMlike conductance. We propose that ybdG encodes a low- abundance MscM- type MS channel, which in cells relieves low levels of membrane tension, obviating the need to activate the major MS channels, MscS and MscL.

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KW - expression

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