An Atypical Phr Peptide Regulates the Developmental Switch Protein RapH

Mirouze, Nicolas; Parashar, Vijay; Baker, Melinda D.; Dubnau, David; Neiditch, Matthew B.

doi:10.1128/jb.05860-11

Cited by 26 publications

(25 citation statements)

References 33 publications

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“…Strikingly, when RapP was overexpressed, added synthetic PhrH pentapeptide (DRNTT) or hexapeptide (TDRNTT) upregulated the expression of P spoIIG -luc. We previ- ously described why we believe the hexapeptide may be the biologically important mature PhrH peptide (28). In contrast, other Phr peptides, including PhrA, PhrC, PhrE, PhrF, and PhrI, as well as the pro-PhrP-derived peptides DRAAT and ADRAAT, had essentially no effect on P spoIIG -luc expression.…”

Section: Resultsmentioning

confidence: 99%

“…phr genes typically encode proPhr polypeptides, which are secreted from the cell, where they undergo proteolytic maturation (25,26). Mature Phr pentapeptides (25,27) or hexapeptides (28) are imported into the cell (29), where each peptide inhibits its cognate Rap protein. For example, PhrA binds to RapA and inhibits its phosphatase activity (30) and PhrF binds to RapF and disrupts the binding of RapF to ComA (23).…”

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confidence: 99%

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A Plasmid-Encoded Phosphatase Regulates Bacillus subtilis Biofilm Architecture, Sporulation, and Genetic Competence

et al. 2013

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bBacillus subtilis biofilm formation is tightly regulated by elaborate signaling pathways. In contrast to domesticated lab strains of B. subtilis which form smooth, essentially featureless colonies, undomesticated strains such as NCIB 3610 form architecturally complex biofilms. NCIB 3610 also contains an 80-kb plasmid absent from laboratory strains, and mutations in a plasmid-encoded homolog of a Rap protein, RapP, caused a hyperrugose biofilm phenotype. Here we explored the role of rapP phrP in biofilm formation. We found that RapP is a phosphatase that dephosphorylates the intermediate response regulator Spo0F. RapP appears to employ a catalytic glutamate to dephosphorylate the Spo0F aspartyl phosphate, and the implications of the RapP catalytic glutamate are discussed. In addition to regulating B. subtilis biofilm formation, we found that RapP regulates sporulation and genetic competence as a result of its ability to dephosphorylate Spo0F. Interestingly, while rap phr gene cassettes routinely form regulatory pairs; i.e., the mature phr gene product inhibits the activity of the rap gene product, the phrP gene product did not inhibit RapP activity in our assays. RapP activity was, however, inhibited by PhrH in vivo but not in vitro. Additional genetic analysis suggests that RapP is directly inhibited by peptide binding. We speculate that PhrH could be subject to posttranslational modification in vivo and directly inhibit RapP activity or, more likely, PhrH upregulates the expression of a peptide that, in turn, directly binds to RapP and inhibits its Spo0F phosphatase activity.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

A Plasmid-Encoded Phosphatase Regulates Bacillus subtilis Biofilm Architecture, Sporulation, and Genetic Competence

et al. 2013

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“…The upper size limits of the peptides that can be imported via the Opp and App oligopeptide ABC transporters of B. subtilis for nutritional purposes are unexplored, but the import of peptides with up to five residues via these systems has been reported (31), and the ligand-binding protein (AppA) of the App transporter can bind nonapeptides (71,72). The Opp and App uptake systems also reimport different Phr-type penta-and hexapeptides excreted by B. subtilis into the growth medium to promote cell-to-cell communication (73) and contribute to the genetic regulation of competence development, sporulation, and extracellular functions in B. subtilis (35)(36)(37)(38)74). Our data now ascribe an additional physiological role to the App, Dpp, Opp, and DtpT peptide transporters by implicating them in the indirect acquisition of the osmoprotectant proline (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…These peptide transport systems (31)(32)(33)(34) allow the acquisition of peptides of various lengths and compositions for use as nutrients and as building blocks for protein synthesis after their proteolytic breakdown. They also enable the import of extracellular peptides as signaling molecules to fine-tune developmental programs leading to competence for DNA uptake and spore formation by B. subtilis (35)(36)(37)(38). Three of these peptide import systems (Dpp, Opp, and App) (31)(32)(33)39) belong to the ABC superfamily of binding-protein-dependent transporters that fuel substrate uptake through ATP hydrolysis.…”

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Osmoprotection of Bacillus subtilis through Import and Proteolysis of Proline-Containing Peptides

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et al. 2013

Appl Environ Microbiol

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Bacillus subtilis can attain cellular protection against the detrimental effects of high osmolarity through osmotically induced de novo synthesis and uptake of the compatible solute L-proline. We have now found that B. subtilis can also exploit exogenously provided proline-containing peptides of various lengths and compositions as osmoprotectants. Osmoprotection by these types of peptides is generally dependent on their import via the peptide transport systems (

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“…Phr peptides originate by posttranslational processing of the product of the phr gene, which is genetically linked and often cotranscribed with the upstream rap gene (24,29,35).…”

Section: T He Initiation Of Sporulation In the Gram-positive Organismmentioning

confidence: 99%

Bacillus subtilis RapA Phosphatase Domain Interaction with Its Substrate, Phosphorylated Spo0F, and Its Inhibitor, the PhrA Peptide

et al. 2012

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Rap proteins in Bacillus subtilis regulate the phosphorylation level or the DNA-binding activity of response regulators such as Spo0F, involved in sporulation initiation, or ComA, regulating competence development. Rap proteins can be inhibited by specific peptides generated by the export-import processing pathway of the Phr proteins. Rap proteins have a modular organization comprising an amino-terminal alpha-helical domain connected to a domain formed by six tetratricopeptide repeats (TPR). In this study, the molecular basis for the specificity of the RapA phosphatase for its substrate, phosphorylated Spo0F (Spo0FϳP), and its inhibitor pentapeptide, PhrA, was analyzed in part by generating chimeric proteins with RapC, which targets the DNAbinding domain of ComA, rather than Spo0FϳP, and is inhibited by the PhrC pentapeptide. In vivo analysis of sporulation efficiency or competence-induced gene expression, as well as in vitro biochemical assays, allowed the identification of the aminoterminal 60 amino acids as sufficient to determine Rap specificity for its substrate and the central TPR3 to TPR5 (TPR3-5) repeats as providing binding specificity toward the Phr peptide inhibitor. The results allowed the prediction and testing of key residues in RapA that are essential for PhrA binding and specificity, thus demonstrating how the widespread structural fold of the TPR is highly versatile, using a common interaction mechanism for a variety of functions in eukaryotic and prokaryotic organisms. T he initiation of sporulation in the Gram-positive organismBacillus subtilis is regulated by the complex phosphorelay signal transduction system. In this system, sporulation-activating signals are sensed by multiple sensor histidine kinases whose activation results in autophosphorylation and phosphoryl transfer to an intermediate component, the Spo0F response regulator. From phosphorylated Spo0F (Spo0FϳP), the phosphoryl group is then transferred to the Spo0B phosphotransferase, which then relays it to the Spo0A response regulator and transcription factor. The phosphorylation level of Spo0A in the cell is the determining factor of whether sporulation will initiate or not in response to the activating signals sensed by the kinases (7,15,22,23).In order to counteract the kinase activities and prevent untimely initiation of sporulation, a number of phosphatases exist to respond to physiological states antithetical to sporulation, such as growth and competence for DNA transformation. There are two families of phosphatases, classified by structure and substrate specificity: the Spo0E-like family and the Rap family (30,32).The members of the Spo0E family of phosphatases (Spo0E, YisI, and YnzD) are small proteins (56 to 85 amino acids) identified by a conserved sequence motif, SQ/RE/DLD, in which the aspartate in the fifth position is the essential catalytic residue.

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An Atypical Phr Peptide Regulates the Developmental Switch Protein RapH

Cited by 26 publications

References 33 publications

A Plasmid-Encoded Phosphatase Regulates Bacillus subtilis Biofilm Architecture, Sporulation, and Genetic Competence

A Plasmid-Encoded Phosphatase Regulates Bacillus subtilis Biofilm Architecture, Sporulation, and Genetic Competence

Osmoprotection of Bacillus subtilis through Import and Proteolysis of Proline-Containing Peptides

Bacillus subtilis RapA Phosphatase Domain Interaction with Its Substrate, Phosphorylated Spo0F, and Its Inhibitor, the PhrA Peptide

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