Signal Peptidase Is Necessary and Sufficient for Site 1 Cleavage of RsiV in Bacillus subtilis in Response to Lysozyme

Castro, Ana N.; Lewerke, Lincoln T.; Hastie, Jessica L.; Ellermeier, Craig D.

doi:10.1128/jb.00663-17

Cited by 20 publications

(36 citation statements)

References 68 publications

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“…Mutation of the cleavage site blocks RsiV degradation and σ V activation (Hastie et al , ). In contrast to other ECF anti‐σ factors, which have dedicated site‐1 proteases, cleavage of RsiV at site‐1 requires signal peptidase (Hastie et al , ; Castro et al , ). B. subtilis encodes five type 1 signal peptidases, four prokaryotic type 1 signal peptidase (SipS, SipT, SipU and SipV) and one eukaryotic type 1 signal peptidase (SipW) (Tjalsma et al , ; Chu et al , ).…”

Section: Degradation Of Rsivmentioning

confidence: 99%

Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Ellermeier

2019

Molecular Microbiology

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σ V is an extracytoplasmic function (ECF) σ factor that is found exclusively in Firmicutes including Bacillus subtilis and the opportunistic pathogens Clostridioides difficile and Enterococcus faecalis. σ V is activated by lysozyme and is required for lysozyme resistance. The activity of σ V is normally inhibited by the anti-σ factor RsiV, a transmembrane protein. RsiV acts as a receptor for lysozyme. The binding of lysozyme to RsiV triggers a signal transduction cascade which results in degradation of RsiV and activation of σ V . Like the anti-σ factors for several other ECF σ factors, RsiV is degraded by a multistep proteolytic cascade that is regulated at the step of site-1 cleavage. Unlike other anti-σ factors, site-1 cleavage of RsiV is not dependent upon a site-1 protease whose activity is regulated. Instead constitutively active signal peptidase cleaves RsiV at site-1 in a lysozyme-dependent manner. The activation of σ V leads to the transcription of genes, which encode proteins required for lysozyme resistance. Extracytoplasmic function (ECF) σ factor backgroundECF σ factors belong to the σ 70 family of σ factors and contain only the σ 2 and σ 4.2 domains. These are homologous to the σ 70 σ 2 and σ 4.2 domains and are required for binding to the -35 and -10 regions of target promoters (Helmann, 2002;2016;Staroń et al., 2009). In addition, most ECF σ factors are required for their own expression. The activity of most ECF σ factors is inhibited by a cognate anti-σ factor which is often encoded within the σ factor operon. Many but not all anti-σ factors are membrane proteins (Staroń et al., 2009). The σ factor activity is induced by inhibiting the function of the anti-σ factor. Activation of the σ factor can occur by one of several mechanisms; a conformational change in the anti-σ factor releases the σ factor, an anti-anti-σ factor binds to the anti-σ factor inducing σ factor release, or destruction of the anti-σ factor via regulated intramembrane proteolysis (Ho and Ellermeier, 2012;Helmann, 2016). Here, we will focus on activation of ECF σ factors by degradation of the anti-σ factor. In the presence of an inducing signal, a site-1 protease cleaves the extracellular domain of an anti-σ factor. Following site-1 protease cleavage, the truncated anti-σ factor is cleaved by a site-2 protease within the transmembrane region of the anti-σ factor. The remainder of the anti-σ factor is then degraded by cytosolic proteases. This frees the σ factor to interact with RNA polymerase and transcribe target genes (Brown et al., 2000;Ho and Ellermeier, 2012;Helmann, 2016).The ECF σ factor σ V is found exclusively in Firmicutes or low GC Gram-positive bacteria and belongs to the ECF30 family of ECF σ factors (Staroń et al., 2009). The σ V system is encoded by the model organism Bacillus subtilis, as well as the opportunistic pathogens Enterococcus faecalis and Clostridioides difficile (Fig. 1). σ V is not present in all Firmicutes or even closely related species. While present in B. subtilis, σ V is not present in B. anthrac...

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Section: Degradation Of Rsivmentioning

confidence: 99%

Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Ellermeier

2019

Molecular Microbiology

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“…SigV is induced by lysozyme and confers lysozyme resistance by O‐acetylation of the PG and D‐alanylation of teichoic acids (Ho et al ., ; Guariglia‐Oropeza and Helmann, ). The activation of SigV begins with the binding of lysozyme to the anti‐sigma factor RsiV, which facilitates proteolysis by signal peptidase cleavage at site 1 (Castro et al , ) followed by intramembrane protease RasP cleavage at site 2 (Hastie et al , ). It is not yet clear how SigM is activated and whether proteolysis is involved.…”

Section: Introductionmentioning

confidence: 99%

Deciphering the essentiality and function of the anti‐σ^M factors in Bacillus subtilis

Zhao

Roistacher

Helmann

2019

Molecular Microbiology

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Bacteria use alternative sigma factors to adapt to different growth and stress conditions. The Bacillus subtilis extracytoplasmic function sigma factor SigM regulates genes for cell wall synthesis and is crucial for maintaining cell wall homeostasis under stress conditions. The activity of SigM is regulated by its anti-sigma factor, YhdL, and the accessory protein YhdK. Here, we show that dysregulation of SigM caused by the absence of either component of the anti-sigma factor complex leads to toxic levels of SigM and severe growth defects. High SigM activity results from a dysregulated positive feedback loop, and can be suppressed by overexpression of the housekeeping sigma, SigA. Using a sigM merodiploid strain, we selected for suppressor mutations that allow survival of yhdL depletion strain. The recovered suppressor mutations map to the beta and beta-prime subunits of RNA polymerase core enzyme and selectively reduce SigM activity, and in some cases increase the activity of other alternative sigma factors. This work highlights the ability of mutations in RNA polymerase that remodel the sigma-core interface to differentially affect sigma factor activity, and thereby alter the transcriptional landscape of the cell.

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“…The anti-σ factors RseA in E. coli as well as RsiW and RsiV in B. subtilis are degraded sequentially by regulated intramembrane proteolysis. Each of these anti-σ factors requires a different family of proteases to cleave the anti-σ factor at site-1 (14, 22, 30, 47, 48) while site-2 cleavage is carried out by the conserved site-2 protease (14, 15, 35). We hypothesize that σ P is activated in a similar manner.…”

Section: Discussionmentioning

confidence: 99%

Activation of the Extracytoplasmic Function σ factor σ^Pby β-lactams inBacillus thuringiensisrequires the site-2 protease RasP

Nauta

Müh

et al. 2019

Preprint

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30Bacteria can utilize alternative σ factors to regulate sets of genes in response to 31 changes in the environment. The largest and most diverse group of alternative σ factors are the 32 Extracytoplasmic Function (ECF) σ factors. σ P is an ECF σ factor found in Bacillus anthracis, B. 33 cereus, and B. thuringiensis. Previous work showed σ P is induced by ampicillin, a β-lactam 34 antibiotic, and required for resistance to ampicillin. However, it was not known how activation of 35 σ P is controlled or what other antibiotics may activate σ P . Here we report that activation of σ P is 36 specific to a subset of β-lactams and σ P is required for resistance to these β-lactams. We 37 demonstrate that activation of σ P is controlled by the proteolytic destruction of the anti-σ factor, 38RsiP, and that degradation of RsiP requires multiple proteases. Upon exposure to β-lactams, 39 the extracellular domain of RsiP is cleaved by an unknown protease, which we predict cleaves 40 at site-1. Following cleavage by the unknown protease, the N-terminus of RsiP is further 41 degraded by the site-2 intramembrane protease, RasP. Our data indicate that RasP cleavage of 42RsiP is not the rate-limiting step in σ P activation. This proteolytic cascade leads to activation of 43 σ P which induces resistance to β-lactams likely via increased expression of β-lactamases. 44 45 Importance 46

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Signal Peptidase Is Necessary and Sufficient for Site 1 Cleavage of RsiV in Bacillus subtilis in Response to Lysozyme

Cited by 20 publications

References 68 publications

Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Deciphering the essentiality and function of the anti‐σ^M factors in Bacillus subtilis

Activation of the Extracytoplasmic Function σ factor σ^Pby β-lactams inBacillus thuringiensisrequires the site-2 protease RasP

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Signal Peptidase Is Necessary and Sufficient for Site 1 Cleavage of RsiV in Bacillus subtilis in Response to Lysozyme

Cited by 20 publications

References 68 publications

Activation of the extracytoplasmic function σ factor σV by lysozyme

Activation of the extracytoplasmic function σ factor σV by lysozyme

Deciphering the essentiality and function of the anti‐σM factors in Bacillus subtilis

Activation of the Extracytoplasmic Function σ factor σPby β-lactams inBacillus thuringiensisrequires the site-2 protease RasP

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Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Activation of the extracytoplasmic function σ factor σ^V by lysozyme

Deciphering the essentiality and function of the anti‐σ^M factors in Bacillus subtilis

Activation of the Extracytoplasmic Function σ factor σ^Pby β-lactams inBacillus thuringiensisrequires the site-2 protease RasP