Identification of subtilisin, Epr and Vpr as enzymes that produce CSF, an extracellular signalling peptide of <i>Bacillus subtilis</i>

Lanigan-Gerdes, Sara; Dooley, Alek N.; Faull, Kym F.; Lazazzera, Beth

doi:10.1111/j.1365-2958.2007.05869.x

Cited by 67 publications

(79 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, Lanigan-Gerdes et al (2007) showed that the PhrC propeptide could be cleaved to release CSF from the extreme C terminus by any one of three secreted proteases. Therefore, our results are consistent with a role in swarming for the residual PhrC 35 aa residue peptide (including the signal sequence), which should be exposed on the exterior of the cytoplasmic membrane.…”

Section: Discussionmentioning

confidence: 99%

“…Subsequent processing by different cell-wall-associated proteases results in the release of the CSF pentapeptide from the C terminus of PhrC (Lanigan-Gerdes et al, 2007). Studies in liquid cultures have clearly demonstrated the presence of CSF in culture supernatants (around 10 nM), and that CSF can be imported by the Opp permease into the cytosol, where it promotes activation of srf expression via competition with RapC (Core & Perego, 2003).…”

Section: Evidence That Phrc Plays a Novel Role In Swarmingmentioning

confidence: 99%

“…This is defined as the transparent region extending from the central mother colony to the visible perimeter of the zone, and is normally detected as spreading just ahead of the swarm front (position indicated by black arrows in the relevant panels in Figs 2a and 3). Notably, comXP (and comQ, not shown), although having substantially reduced levels of expression of srf (lacZ fusion data) in liquid cultures (Hahn & Dubnau, 1991;Cosby et al, 1998;Schneider et al, 2002;Lanigan-Gerdes et al, 2007), produced large surfactin zones. Overall, these results highlighted apparently important differences between liquid cultures and cells on swarm plates, with respect to the regulation of srf expression.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Identification of genes required for different stages of dendritic swarming in Bacillus subtilis, with a novel role for phrC

et al. 2009

View full text Add to dashboard Cite

Highly branched dendritic swarming of B. subtilis on synthetic B-medium involves a developmental-like process that is absolutely dependent on flagella and surfactin secretion. In order to identify new swarming genes, we targeted the two-component ComPA signalling pathway and associated global regulators. In liquid cultures, the histidine kinase ComP, and the response regulator ComA, respond to secreted pheromones ComX and CSF (encoded by phrC) in order to control production of surfactin synthases and ComS (competence regulator). In this study, for what is believed to be the first time, we established that distinct early stages of dendritic swarming can be clearly defined, and that they are amenable to genetic analysis. In a mutational analysis producing several mutants with distinctive phenotypes, we were able to assign the genes sfp (activation of surfactin synthases), comA, abrB and codY (global regulators), hag (flagellin), mecA and yvzB (hag-like), and swrB (motility), to the different swarming stages. Surprisingly, mutations in genes comPX, comQ, comS, rapC and oppD, which are normally indispensable for import of CSF, had only modest effects, if any, on swarming and surfactin production. Therefore, during dendritic swarming, surfactin synthesis is apparently subject to novel regulation that is largely independent of the ComXP pathway; we discuss possible alternative mechanisms for driving srfABCD transcription. We showed that the phrC mutant, largely independent of any effect on surfactin production, was also, nevertheless, blocked early in swarming, forming stunted dendrites, with abnormal dendrite initiation morphology. In a mixed swarm co-inoculated with phrC sfp + and phrC + sfp (GFP), an apparently normal swarm was produced. In fact, while initiation of all dendrites was of the abnormal phrC type, these were predominantly populated by sfp cells, which migrated faster than the phrC cells. This and other results indicated a specific migration defect in the phrC mutant that could not be trans-complemented by CSF in a mixed swarm. CSF is the C-terminal pentapeptide of the surface-exposed PhrC pre-peptide and we propose that the residual PhrC 35 aa residue peptide anchored in the exterior of the cytoplasmic membrane has an apparently novel extracellular role in swarming.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Evidence That Phrc Plays a Novel Role In Swarmingmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Identification of genes required for different stages of dendritic swarming in Bacillus subtilis, with a novel role for phrC

et al. 2009

View full text Add to dashboard Cite

show abstract

“…Interestingly, Vpr was identified as one of the most abundant proteins in biofilms produced by some B. subtilis cells (71). Moreover, this minor serine protease was demonstrated to play a role in the processing of the peptide antibiotic subtilin (72) and production of two quorum sensing signaling peptides, PhrA and CSF, by cleavage of their precursors (73).…”

Section: Discussionmentioning

confidence: 99%

CodY Regulates Expression of the Bacillus subtilis Extracellular Proteases Vpr and Mpr

et al. 2015

View full text Add to dashboard Cite

CodY is a global transcriptional regulator in low-G؉C Gram-positive bacteria that is responsive to GTP and branched-chain amino acids. By interacting with its two cofactors, it is able to sense the nutritional and energetic status of the cell and respond by regulating expression of adaptive genetic programs. In Bacillus subtilis, more than 200 genes, including those for peptide transporters, intracellular proteolytic enzymes, and amino acid degradative pathways, are controlled by CodY. In this study, we demonstrated that expression of two extracellular proteases, Vpr and Mpr, is negatively controlled by CodY. By gel mobility shift and DNase I footprinting assays, we showed that CodY binds to the regulatory regions of both genes, in the vicinity of their transcription start points. The mpr gene is also characterized by the presence of a second, higher-affinity CodY-binding site located at the beginning of its coding sequence. Using strains carrying vpr-or mpr-lacZ transcriptional fusions in which CodY-binding sites were mutated, we demonstrated that repression of both protease genes is due to the direct effect by CodY and that the mpr internal site is required for regulation. The vpr promoter is a rare example of a sigma H-dependent promoter that is regulated by CodY. In a codY null mutant, Vpr became one of the more abundant proteins of the B. subtilis exoproteome. IMPORTANCE CodY is a global transcriptional regulator of metabolism and virulence in low-G؉C

show abstract

“…Minor extracellular protease Vpr has been reported in B. subtilis (Park et al, 2002;Sloma et al, 1991) and Bacillus licheniformis (Ageitos et al, 2007), and it has been shown that Vpr has major roles in subtilisin (lantibiotic) (Corvey et al, 2003) processing, competence and sporulation factor (CSF) processing, and probably in biofilm formation and quorum sensing (Lanigan-Gerdes et al, 2007). Moreover in B. subtilis, Vpr was identified as a fibrinolytic enzyme (Park et al, 2002) and in B. licheniformis, Vpr was reported as one of the milk-clotting proteases (Ageitos et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Cloning of feather-degrading minor extracellular protease from Bacillus cereus DCUW: dissection of the structural domains

2009

View full text Add to dashboard Cite

Bacterial extracellular proteases play an important role in cell survival and cell-cell communication. A high-molecular-mass minor extracellular protease (Vpr) from a featherdegrading bacterium, Bacillus cereus DCUW, has been reported by our laboratory. In the present study, we cloned and expressed Vpr in Escherichia coli. Complete nucleotide sequencing of this gene predicted that the protease is a member of the serine protease family, and SMART domain analysis revealed that the protease consists of an N-terminal signal sequence for secretion, a subtilisin_N sequence that is a signature for N-terminal processing, a catalytic S_8 peptidase domain, and finally a long C-terminal protease-associated (PA) region containing nine intrinsically disordered subdomains. Four truncated constructs of the Vpr protease were cloned and expressed in E. coli. We found that the catalytic domain (amino acid residues 172-583) is sufficient for protease activity. Maturation of the Vpr protease needed both N-terminal and Cterminal processing. We have demonstrated that the oligomerization property is associated with the C-terminal protease-associated domain and also shown that the substrate-binding specificity to raw feather resides in this domain.

show abstract

Identification of subtilisin, Epr and Vpr as enzymes that produce CSF, an extracellular signalling peptide of Bacillus subtilis

Cited by 67 publications

References 55 publications

Identification of genes required for different stages of dendritic swarming in Bacillus subtilis, with a novel role for phrC

Identification of genes required for different stages of dendritic swarming in Bacillus subtilis, with a novel role for phrC

CodY Regulates Expression of the Bacillus subtilis Extracellular Proteases Vpr and Mpr

Cloning of feather-degrading minor extracellular protease from Bacillus cereus DCUW: dissection of the structural domains

Contact Info

Product

Resources

About