Molecular analysis of an operon in Bacillus subtilis encoding a novel ABC transporter with a role in exoprotein production, sporulation and competence

Leskelä, Soile; Kontinen, Vesa P.; Sarvas, Matti

doi:10.1099/13500872-142-1-71

Cited by 40 publications

(44 citation statements)

References 33 publications

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“…Three of them, sup-6, sup-15, and sup-16, displayed the largest increase of secretion. The presence of the ecsA26 mutation in these suppressor mutants was confirmed by demonstrating the presence of an ecsA26 mutation-specific XbaI restriction site in a PCR fragment amplified from ecsA26 DNA (14).…”

Section: Resultsmentioning

confidence: 91%

“…We have found in B. subtilis an ATP-binding cassette (ABC) transporter, designated Ecs, which influences the production of extracellular proteins, competence development, and sporulation (14,15). Accumulation of precursors of secretory proteins in mutant cells with a defective ATPase subunit, EcsA, or integral membrane component, EcsB, suggested that Ecs is a regulator of protein secretion (15).…”

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confidence: 99%

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ClpXP Protease Regulates the Signal Peptide Cleavage of Secretory Preproteins inBacillus subtiliswith a Mechanism Distinct from That of the Ecs ABC Transporter

Pummi¹,

Leskelä²,

Wahlström³

et al. 2002

J Bacteriol

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Identification and characterization of a suppressor mutation, sup-15, which partially restored secretion in the protein secretion-deficient Bacillus subtilis ecsA26 mutant, led us to discover a novel function of Clp protease. Inactivation of ClpP improved the processing of the precursor of AmyQ ␣-amylase exposed on the outer surface of the cytoplasmic membrane. A similar improvement of AmyQ secretion was conferred by inactivation of the ClpX substrate-binding component of the ClpXP complex. In the absence of ClpXP, the transcription of the sipS, sipT, sipV, and lsp signal peptidase genes was elevated two-to fivefold, a likely cause of the improvement of the processing and secretion of AmyQ and complementation of ecs mutations. Specific overproduction of SipT enhanced the secretion. These findings extend the regulatory roles of ClpXP to protein secretion. ClpXP also influenced the processing of the lipoprotein PrsA. A concerted regulation of signal peptidase genes by a ClpXP-dependent activator is suggested. In contrast, Ecs did not affect transcription of the sip genes, pointing to a different mechanism of secretion regulation.

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Section: Resultsmentioning

confidence: 91%

mentioning

confidence: 99%

ClpXP Protease Regulates the Signal Peptide Cleavage of Secretory Preproteins inBacillus subtiliswith a Mechanism Distinct from That of the Ecs ABC Transporter

Pummi¹,

Leskelä²,

Wahlström³

et al. 2002

J Bacteriol

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“…The ecsA gene encodes the ATPase subunit and ecsB encodes the transmembrane subunit of an ABC transporter referred to as Ecs (15,21). The ecsC gene encodes a protein that shows no significant sequence similarity to any previously described ABC transporter component or accessory protein or indeed to any protein of known function.…”

Section: Vol 186 2004 Genetic Analysis Of B Subtilis Community Devmentioning

confidence: 99%

Genes Involved in Formation of Structured Multicellular Communities byBacillus subtilis

et al. 2004

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The spore-forming bacterium Bacillus subtilis is capable of assembling multicellular communities (biofilms) that display a high degree of spatiotemporal organization. Wild strains that have not undergone domestication in the laboratory produce particularly robust biofilms with complex architectural features, such as fruitingbody-like aerial projections whose tips serve as preferential sites for sporulation. To discover genes involved in this multicellular behavior and to do so on a genome-wide basis, we took advantage of a large collection of mutants which have disruptions of most of the uncharacterized genes in the B. subtilis genome. This collection, which was generated with a laboratory strain, was screened for mutants that were impaired in biofilm formation. This subset of mutated genes was then introduced into the wild strain NCIB 3610 to study their effects on biofilm formation in liquid and solid media. In this way we identified six genes that are involved in the development of multicellular communities. These are yhxB (encoding a putative phosphohexomutase that may mediate exopolysaccharide synthesis), sipW (encoding a signal peptidase), ecsB (encoding an ABC transporter subunit), yqeK (encoding a putative phosphatase), ylbF (encoding a regulatory protein), and ymcA (a gene of unknown function). Further analysis revealed that these six genes play different roles in B. subtilis community development.Formation of biofilms-surface-associated multicellular assemblages-is an important microbial survival strategy (4, 25). Relative to the rapid, continual, and extreme changes in environmental conditions that can characterize a planktonic existence, biofilms offer the constituent cells some shelter, enabling them to establish long-term relationships with each other and their immediate surroundings. Cells in different regions of a developing biofilm experience diverse environmental conditions, resulting in marked patterns of cellular differentiation (e.g., see references 5 and 16). Such spatiotemporal organization is particularly striking in biofilms formed by wild strains of the spore-forming bacterium Bacillus subtilis. Unlike strains that have been domesticated by decades of propagation in the laboratory (e.g., B. subtilis 168), which form thin and relatively undifferentiated biofilms, wild B. subtilis strains form elaborate multicellular communities that display conspicuous architectural features, such as fruiting-body-like aerial projections that extend from the surfaces of the biofilm. The tips of these fruiting bodies serve as preferential sites for spore formation (2).We are interested in discovering genes that are involved in biofilm and fruiting body formation. We have focused our efforts on a relatively undomesticated strain of B. subtilis, NCIB 3610 (hereafter referred to as "3610"), which forms robust and highly structured biofilms both in liquid and on solid medium (colonies). When inoculated into a standing culture of minimal medium, 3610 initially grows planktonically as motile, single cells. The ...

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“…Different transposon insertions in ecsA or ecsB causing stabilization of the GFP-RsiW reporter and abolishing induction of a transcriptional fusion of lacZ to the strong s W -controlled yuaF promoter were obtained. These genes encode the ABC and the transmembrane part of an ABC transporter that has been characterized previously (Leskela et al, 1996), orthologues of which can be found in a great variety of bacterial species. However, the function of EcsAB is still unknown.…”

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The Bacillus subtilis ABC transporter EcsAB influences intramembrane proteolysis through RasP

et al. 2008

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The Bacillus subtilis s W regulon is induced by different stresses that most probably affect integrity of the cell envelope. The activity of the extracytoplasmic function (ECF) sigma factor s W is modulated by the transmembrane anti-sigma factor RsiW, which undergoes stress-induced degradation in a process known as regulated intramembrane proteolysis, finally resulting in the release of s W and the transcription of s W -controlled genes. Mutations in the ecsA gene, which encodes an ATP binding cassette (ABC) of an ABC transporter of unknown function, block site-2 proteolysis of RsiW by the intramembrane cleaving protease RasP (YluC). In addition, degradation of the cell division protein FtsL, which represents a second RasP substrate, is blocked in an ecsA-negative strain. The defect in s W induction of an ecsA-knockout strain could be partly suppressed by overproducing RasP. A B. subtilis rasP-knockout strain displayed the same pleiotropic phenotype as an ecsA knockout, namely defects in processing a-amylase, in competence development, and in formation of multicellular structures known as biofilms. INTRODUCTIONGenes of Bacillus subtilis controlled by the alternative extracytoplasmic function (ECF) sigma factor s W constitute an antibiosis regulon (Helmann, 2002(Helmann, , 2006 that responds to a variety of envelope stresses such as certain antibiotics (Cao et al., 2002) and antimicrobial peptides (Pietiäinen et al., 2005;Butcher & Helmann, 2006), and also alkaline shock and phage infection . Its activity is modulated by RsiW, a transmembrane anti-sigma factor that sequesters and inactivates s W . Upon a stress signal, RsiW is degraded by the mechanism of regulated intramembrane proteolysis (RIP). In a concerted action, at least three proteases in three different compartments of the cell degrade the RsiW anti-sigma factor in a sequential manner, finally resulting in the release of s W and the transcription of s Wcontrolled genes. The first protease that has been identified to be involved in that process is RasP (regulating anti-sigma factor protease; formerly YluC). RasP belongs to the group of zinc-dependent intramembrane cleaving proteases (iClips) and cleaves RsiW in its transmembrane domain after the extracytoplasmic part of the anti-sigma factor has been removed by a site-1 protease (Schöbel et al., 2004). The site-2 degradation step catalysed by RasP uncovers a cryptic proteolytic tag that ensures further degradation of the RsiW remnant by cytoplasmic proteases, mainly ClpXP (Zellmeier et al., 2006). These two steps are related to RIP of the s E anti-sigma factor RseA of Escherichia coli (Ades, 2004;Alba & Gross, 2004). RasP is an orthologue of E. coli RseP, and the concept of a cryptic proteolytic tag recognized by ClpXP was first described for that system (Akiyama et al., 2004;Flynn et al., 2004). However, there is a marked difference in the site-1 proteolytic step, because no dependency on B. subtilis Deg (Htr) proteases was found and the inducing stress is apparently different. The probable site-1 prot...

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Molecular analysis of an operon in Bacillus subtilis encoding a novel ABC transporter with a role in exoprotein production, sporulation and competence

Cited by 40 publications

References 33 publications

ClpXP Protease Regulates the Signal Peptide Cleavage of Secretory Preproteins inBacillus subtiliswith a Mechanism Distinct from That of the Ecs ABC Transporter

ClpXP Protease Regulates the Signal Peptide Cleavage of Secretory Preproteins inBacillus subtiliswith a Mechanism Distinct from That of the Ecs ABC Transporter

Genes Involved in Formation of Structured Multicellular Communities byBacillus subtilis

The Bacillus subtilis ABC transporter EcsAB influences intramembrane proteolysis through RasP

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