SwrA regulates assembly of Bacillus subtilis DegU via its interaction with N-terminal domain of DegU

Ogura, Mitsuo; Tsukahara, Kensuke

doi:10.1093/jb/mvs036

Cited by 40 publications

(32 citation statements)

References 42 publications

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“…The degU gene resides within an operon with the upstream degS gene (14,15). DegU, a DNA-binding protein that recognizes AT-rich octamers in a variety of arrangements (12,16), controls many genes and biological processes (17,18). In its low-level phosphorylation state, including its nonphosphorylated state, DegU activates comK (a master regulator of genetic competence) and the fla/che operon (12,14,19,20).…”

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The Bacillus subtilis Response Regulator Gene degU Is Positively Regulated by CcpA and by Catabolite-Repressed Synthesis of ClpC

2013

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In Bacillus subtilis, the response regulator DegU and its cognate kinase, DegS, constitute a two-component system that regulates many cellular processes, including exoprotease production and genetic competence. Phosphorylated DegU (DegU-P) activates its own promoter and is degraded by the ClpCP protease. We observed induction of degU by glucose in sporulation medium. This was abolished in two mutants: the ccpA (catabolite control protein A) and clpC disruptants. Transcription of the promoter of the operon containing clpC (PclpC) decreased in the presence of glucose, and the disruption of ccpA resulted in derepression of PclpC. However, this was not directly mediated by CcpA, because we failed to detect binding of CcpA to PclpC. Glucose decreased the expression of clpC, leading to low cellular concentrations of the ClpCP protease. Thus, degU is induced through activation of autoregulation by a decrease in ClpCP-dependent proteolysis of DegU-P. An electrophoretic mobility shift assay showed that CcpA bound directly to the degU upstream region, indicating that CcpA activates degU through binding. The bound region was narrowed down to 27 bases, which contained a cre (catabolite-responsive element) sequence with a low match to the cre consensus sequence. In a footprint analysis, CcpA specifically protected a region containing the cre sequence from DNase I digestion. The induction of degU by glucose showed complex regulation of the degU gene.

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“…The other protein-protein interaction includes the small protein SwrA, which is a swarming motility regulator (32). SwrA interacts with the N-terminal region of DegU, leading to stabilization of the DegU dimer (16).…”

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The Bacillus subtilis Response Regulator Gene degU Is Positively Regulated by CcpA and by Catabolite-Repressed Synthesis of ClpC

2013

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“…However, this gene was recently shown to be dispensable for swarming on synthetic B medium under high-humidity conditions (14). Through the action of the transcription-activating protein DegU (30), SwrA stimulates the transcription of the fla-che operon, which contains the majority of genes for flagellar and chemotaxis proteins and sigD, the Dencoding gene (21,32). This increases the synthesis of structural flagellar proteins, including flagellin, and stimulates the expression of swrA itself through a D -dependent promoter (5).…”

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Contribution of Surfactin and SwrA to Flagellin Expression, Swimming, and Surface Motility in Bacillus subtilis

Ghelardi¹,

Salvetti²,

Ceragioli

et al. 2012

Appl Environ Microbiol

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ABSTRACTMulticellular communities produced byBacillus subtiliscan adopt sliding or swarming to translocate over surfaces. While sliding is a flagellum-independent motility produced by the expansive forces in a growing colony, swarming requires flagellar functionality and is characterized by the appearance of hyperflagellated swarm cells that associate in bundles or rafts during movement. Previous work has shown that swarming by undomesticatedB. subtilisstrains requiresswrA, a gene that upregulates the expression of flagellar genes and increases swimming motility, and surfactin, a lipopeptide biosurfactant that also facilitates sliding. Through an analysis ofswrA+andswrAmutant laboratory strains with or without a mutation insfp(a gene involved in surfactin production), we show that bothswrAand surfactin upregulate the transcription of the flagellin gene and increase bacterial swimming. Surfactin also allows the nonswarmingswrAmutant strain to efficiently colonize moist surfaces by sliding. Finally, we reconfirm the essential role ofswrAin swarming and show that surfactin, which increases surface wettability, allowsswrA+strains to produce swarm cells on media at low humidity.

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“…The His tag was removed from His-SinR by biotinylated thrombin protease. His-DegS and intact DegU were purified from E. coli BL21(DE3) carrying pET-degS and pTYB-degU, as described previously (10,34). DegU was dialyzed against MOPS (morpholinepropanesulfonic acid)-based degradation assay buffer (35).…”

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Regulation of the Response Regulator Gene degU through the Binding of SinR/SlrR and Exclusion of SinR/SlrR by DegU in Bacillus subtilis

2014

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bBacillus subtilis DegU is a response regulator of the DegS-DegU two-component regulatory system. Phosphorylated DegU (DegU-P) controls many genes and biological processes, such as exoprotease and ␥-polyglutamic acid production, in addition to the degU gene, by binding to target gene promoters. Nonphosphorylated DegU and low levels of DegU-P are required for swarming motility and genetic competence. The DNA-binding repressors SinR and SlrR are part of a double-negative feedback loop and comprise the epigenetic switch governing biofilm formation. In this study, we found that SinR repressed degU. Furthermore, SlrR, which interacts with SinR through protein-protein interaction, seems to have an active role in degU expression in in vivo lacZ analysis. An in vitro transcription assay supported this observation. An electrophoretic mobility shift assay (EMSA) showed that SinR bound to the degU promoter and that SlrR formed a complex with SinR on the degU promoter. In EMSA, DegU-P excluded the SinR/SlrR complex but not SinR from the degU promoter in the presence of RNA polymerase. These findings suggest that DegU-P interacts with SlrR. In support of this hypothesis, disruption of the slrR gene resulted in decreased degU expression. This newly identified regulatory mechanism for degU is considered to be sequential transcription factor replacement.

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SwrA regulates assembly of Bacillus subtilis DegU via its interaction with N-terminal domain of DegU

Cited by 40 publications

References 42 publications

The Bacillus subtilis Response Regulator Gene degU Is Positively Regulated by CcpA and by Catabolite-Repressed Synthesis of ClpC

The Bacillus subtilis Response Regulator Gene degU Is Positively Regulated by CcpA and by Catabolite-Repressed Synthesis of ClpC

Contribution of Surfactin and SwrA to Flagellin Expression, Swimming, and Surface Motility in Bacillus subtilis

Regulation of the Response Regulator Gene degU through the Binding of SinR/SlrR and Exclusion of SinR/SlrR by DegU in Bacillus subtilis

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