Characterization of the sppA gene coding for protease IV, a signal peptide peptidase of Escherichia coli

Suzuki, T; Itoh, Ayako; Ichihara, Shigeyuki; Mizushima, Shôji

doi:10.1128/jb.169.6.2523-2528.1987

Cited by 55 publications

(39 citation statements)

References 37 publications

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Section: Resultsmentioning

confidence: 99%

“…The sppA gene codes for protease IV, a signal peptide peptidase found in the cytoplasmic membrane that digests cleaved signal peptides (13,36). The sppA gene is also not essential for bacterial growth (36). Bacteria which carry a deletion of the gene digest signal peptides at a considerably slower rate than sppA+ bacteria, although this digestion is still significant (36).…”

Section: Resultsmentioning

confidence: 99%

“…The sppA gene is also not essential for bacterial growth (36). Bacteria which carry a deletion of the gene digest signal peptides at a considerably slower rate than sppA+ bacteria, although this digestion is still significant (36). This observation suggests that there is another protease(s) involved in signal peptide digestion in the cell envelope.…”

Section: Resultsmentioning

confidence: 99%

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Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype

et al. 1991

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We cloned and sequenced the sohB gene of Escherichia coli. The temperature-sensitive phenotype of bacteria that carry a Tn10 insertion in the htrA (degP) gene is relieved when the sohB gene is present in the cell on a multicopy plasmid (30 to 50 copies per cell). The htrA gene encodes a periplasmic protease required for bacterial viability only at high temperature, i.e., above 39 degrees C. The sohB gene maps to 28 min on the E. coli chromosome, precisely between the topA and btuR genes. The gene encodes a 39,000-Mr precursor protein which is processed to a 37,000-Mr mature form. Sequencing of a DNA fragment containing the gene revealed an open reading frame which could encode a protein of Mr 39,474 with a predicted signal sequence cleavage site between amino acids 22 and 23. Cleavage at this site would reduce the size of the processed protein to 37,474 Mr. The predicted protein encoded by the open reading frame has homology with the inner membrane enzyme protease IV of E. coli, which digests cleaved signal peptides. Therefore, it is possible that the sohB gene encodes a previously undiscovered periplasmic protease in E. coli that, when overexpressed, can partially compensate for the missing HtrA protein function.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype

et al. 1991

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“…For prokaryotes in general, little is known about the removal of signal peptides from the membrane. E. coli SppA (protease IV) has been shown to degrade the processed signal sequence of the major lipoprotein, but it is not the only protease that is responsible for signal peptide digestion in the cell envelope (Suzuki et al, 1987). For B. subtilis, three proteins similar to SppA have been described (TepA, SppA and YqeZ; Bolhuis et al, 1999;Helmann, 2002).…”

Section: Discussionmentioning

confidence: 99%

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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“…Earlier studies showed that a membrane-bound protease SppA degraded SP from Lpp in detergent extracts (15,16). However, its in vivo function has not been established.…”

Section: Rsep Cleavage Of Sp Requires a Preceding Processing Of Preprmentioning

confidence: 99%

Post-liberation cleavage of signal peptides is catalyzed by the site-2 protease (S2P) in bacteria

Saito

Hizukuri

Matsuo

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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A signal peptide (SP) is cleaved off from presecretory proteins by signal peptidase during or immediately after insertion into the membrane. In metazoan cells, the cleaved SP then receives proteolysis by signal peptide peptidase, an intramembrane-cleaving protease (I-CLiP). However, bacteria lack any signal peptide peptidase member I-CLiP, and little is known about the metabolic fate of bacterial SPs. Here we show that Escherichia coli RseP, an site-2 protease (S2P) family I-CLiP, introduces a cleavage into SPs after their signal peptidase-mediated liberation from preproteins. A Bacillus subtilis S2P protease, RasP, is also shown to be involved in SP cleavage. These results uncover a physiological role of bacterial S2P proteases and update the basic knowledge about the fate of signal peptides in bacterial cells. SPP belongs to intramembrane-cleaving proteases (I-CLiPs), which are classified into SPP/γ-secretase (aspartyl proteases), rhomboid (serine protease), and S2P (zinc metalloprotease) (5, 6). These proteases liberate otherwise membrane-tethered domains of membrane proteins to function as a regulatory molecule. I-CLiPs have specific intramolecular routes that make water molecules accessible to the intramembrane proteolytic active sites (5, 6). For such proteolysis to occur, γ-secretase/SPP and S2P require, in most cases, removal of the ectodomain of the substrate by other protease (5, 7). SPP and S2P prefer substrate with the type II transmembrane orientation and γ-secretase and rhomboid prefer the type I orientation (7).Bacteria contain S2Ps and rhomboids but not SPPs, and only limited knowledge is available about the fate of bacterial SPs (7). We and others have been characterizing RseP, an Escherichia coli member of the S2P involved in the σ E pathway extracytoplasmic stress response (8, 9). In this regulation, a protease, DegS, responds to misassembled outer membrane proteins and introduces the first proteolytic cleavage into RseA, a membraneintegrated anti-σ E protein (10). Subsequently, RseP introduces the second cleavage into RseA, activating σ E to transcribe stressinducible genes (8, 9). Although RseA is the only physiological substrate of RseP so far established, we have shown previously that RseP can cleave a wider range of transmembrane sequences having helix-destabilizing residues (11). Our preliminary results that RseP cleaved a β-lactamase (Bla) fusion protein at or around its SP (11), together with the fact that RseP and SPP share the substrate preference for the type II orientation, prompted us to undertake the present study. Our results suggest strongly that S2Ps (E. coli RseP and Bacillus subtilis RasP) are involved in degradation of remnant SPs left in the bacterial cytoplasmic membrane, in contrast with the currently prevailing concept that SppA (12, 13), a protease unrelated to the S2P or the SPP family, is the enzyme responsible for signal peptide cleavage in bacteria.

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Characterization of the sppA gene coding for protease IV, a signal peptide peptidase of Escherichia coli

Cited by 55 publications

References 37 publications

Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype

Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype

The Bacillus subtilis ABC transporter EcsAB influences intramembrane proteolysis through RasP

Post-liberation cleavage of signal peptides is catalyzed by the site-2 protease (S2P) in bacteria

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