Bacterial sensing: A putative amphipathic helix in RsiV is the switch for activating σV in response to lysozyme

PLoS Genetics
Lincoln T LewerkeCraig D Ellermeier

Abstract

Extra Cytoplasmic Function (ECF) σ factors are a diverse group of alternate σ factors bacteria use to respond to changes in the environment. The Bacillus subtilis ECF σ factor σV responds to lysozyme. In the absence of lysozyme, σV is held inactive by the anti-σ factor, RsiV. In the presence of lysozyme RsiV is degraded via regulated intramembrane proteolysis, which results in the release of σV and thus activation of lysozyme resistance genes. Signal peptidase is required to initiate degradation of RsiV. Previous work indicated that RsiV only becomes sensitive to signal peptidase upon direct binding to lysozyme. We have identified a unique domain of RsiV that is responsible for protecting RsiV from cleavage by signal peptidase in the absence of lysozyme. We provide evidence that this domain contains putative amphipathic helices. Disruption of the hydrophobic surface of these helices by introducing positively charged residues results in constitutive cleavage of RsiV by signal peptidase and thus constitutive σV activation. We provide further evidence that this domain contains amphipathic helices using a membrane-impermeable reagent. Finally, we show that upon lysozyme binding to RsiV, the hydrophobic face of the amphipathic helix...Continue Reading

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Citations

Jul 10, 2019·Molecular Microbiology·Theresa D Ho, Craig D Ellermeier
Feb 12, 2020·Journal of Bacteriology·Stephanie A RaglandAlison K Criss
Nov 17, 2020·Frontiers in Molecular Biosciences·Sarah Wettstadt, María A Llamas
Feb 3, 2021·Journal of Bacteriology·Angela Saadat, Stephen B Melville
Jul 22, 2021·Molecular Systems Biology·Christian P SchwallJames C W Locke
Aug 10, 2021·Journal of Bacteriology·Srivatsan ParthasarathyLynn E Hancock

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Methods Mentioned

BETA
environmental stresses
RIP
Circular Dichroism
PCR

Software Mentioned

- FOLD -
PEP
NetWheels
FOLD3
Clustal Omega

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