Export of the Pseudopilin XcpT of the
            <i>Pseudomonas aeruginosa</i>
            Type II Secretion System via the Signal Recognition Particle-Sec Pathway

Arts, Jorik; Boxtel, Ria van; Filloux, Alain; Tommassen, Jan; Koster, Margot

doi:10.1128/jb.01236-06

Cited by 58 publications

(60 citation statements)

References 55 publications

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“…A defect in prepilin peptidase PulO-mediated processing of PulG was also demonstrated in SRP-depleted strains by using a variant of prePulG in which the propeptide was artificially extended with an N-terminal FLAG tag (data not shown). In the accompanying paper, Arts et al show that processing of native XcpT, a close homologue of PulG, by prepilin peptidase also depends on SRP and the Sec machinery (3).…”

Section: Discussionmentioning

confidence: 99%

Signal Recognition Particle-Dependent Inner Membrane Targeting of the PulG Pseudopilin Component of a Type II Secretion System

et al. 2007

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The pseudopilin PulG is an essential component of the pullulanase-specific type II secretion system from Klebsiella oxytoca. PulG is the major subunit of a short, thin-filament pseudopilus, which presumably elongates and retracts in the periplasm, acting as a dynamic piston to promote pullulanase secretion. It has a signal sequence-like N-terminal segment that, according to studies with green and red fluorescent protein chimeras, anchors unassembled PulG in the inner membrane. We analyzed the early steps of PulG inner membrane targeting and insertion in Escherichia coli derivatives defective in different protein targeting and export factors. The ␤-galactosidase activity in strains producing a PulG-LacZ hybrid protein increased substantially when the dsbA, dsbB, or all sec genes tested except secB were compromised by mutations. To facilitate analysis of native PulG membrane insertion, a leader peptidase cleavage site was engineered downstream from the N-terminal transmembrane segment (PrePulG*). Unprocessed PrePulG* was detected in strains carrying mutations in secA, secY, secE, and secD genes, including some novel alleles of secY and secD. Furthermore, depletion of the Ffh component of the signal recognition particle (SRP) completely abolished PrePulG* processing, without affecting the Sec-dependent export of periplasmic MalE and RbsB proteins. Thus, PulG is cotranslationally targeted to the inner membrane Sec translocase by SRP.

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Section: Discussionmentioning

confidence: 99%

Signal Recognition Particle-Dependent Inner Membrane Targeting of the PulG Pseudopilin Component of a Type II Secretion System

et al. 2007

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“…In contrast, proteins with the unique type III signal sequence are called prepilins and are not competent for assembly until the signal is cleaved at the cytoplasmic face of the membrane (369). Prepilins are inserted into the cytoplasmic membrane by the Sec system (22,144), and the signal sequence is removed by dedicated aspartyl proteases called prepilin peptidases (301). The positively charged signal sequence is likely important for the correct orientation of pilin proteins, with their C-terminal domains outside the cytoplasmic membrane.…”

Section: The Signature Type III Signal Sequencementioning

confidence: 99%

Type IV Pilin Proteins: Versatile Molecular Modules

Giltner

Nguyen

Burrows

2012

Microbiol Mol Biol Rev

410

519

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SUMMARYType IV pili (T4P) are multifunctional protein fibers produced on the surfaces of a wide variety of bacteria and archaea. The major subunit of T4P is the type IV pilin, and structurally related proteins are found as components of the type II secretion (T2S) system, where they are called pseudopilins; of DNA uptake/competence systems in both Gram-negative and Gram-positive species; and of flagella, pili, and sugar-binding systems in the archaea. This broad distribution of a single protein family implies both a common evolutionary origin and a highly adaptable functional plan. The type IV pilin is a remarkably versatile architectural module that has been adopted widely for a variety of functions, including motility, attachment to chemically diverse surfaces, electrical conductance, acquisition of DNA, and secretion of a broad range of structurally distinct protein substrates. In this review, we consider recent advances in this research area, from structural revelations to insights into diversity, posttranslational modifications, regulation, and function.

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“…Proteinase K accessibility experiments were performed as described (Arts et al, 2007). Briefly, P. aeruginosa PAO1 cells were grown to late-exponential phase at 37 uC in DCAA medium.…”

Section: D30pvdamentioning

confidence: 99%

Membrane-association determinants of the ω-amino acid monooxygenase PvdA, a pyoverdine biosynthetic enzyme from Pseudomonas aeruginosa

et al. 2008

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The L-ornithine N d -oxygenase PvdA catalyses the N d -hydroxylation of L-ornithine in many Pseudomonas spp., and thus provides an essential enzymic function in the biogenesis of the pyoverdine siderophore. Here, we report a detailed analysis of the membrane topology of the PvdA enzyme from the bacterial pathogen Pseudomonas aeruginosa. Membrane topogenic determinants of PvdA were identified by computational analysis, and verified in Escherichia coli by constructing a series of translational fusions between PvdA and the PhoA (alkaline phosphatase) reporter enzyme. The inferred topological model resembled a eukaryotic reverse signal-anchor (type III) protein, with a single N-terminal domain anchored to the inner membrane, and the bulk of the protein spanning the cytosol. According to this model, the predicted transmembrane region should overlap the putative FAD-binding site. Cell fractionation and proteinase K accessibility experiments in P. aeruginosa confirmed the membrane-bound nature of PvdA, but excluded the transmembrane topology of its N-terminal hydrophobic region. Mutational analysis of PvdA, and complementation assays in a P. aeruginosa DpvdA mutant, demonstrated the dual (structural and functional) role of the PvdA N-terminal domain.

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Export of the Pseudopilin XcpT of the Pseudomonas aeruginosa Type II Secretion System via the Signal Recognition Particle-Sec Pathway

Cited by 58 publications

References 55 publications

Signal Recognition Particle-Dependent Inner Membrane Targeting of the PulG Pseudopilin Component of a Type II Secretion System

Signal Recognition Particle-Dependent Inner Membrane Targeting of the PulG Pseudopilin Component of a Type II Secretion System

Type IV Pilin Proteins: Versatile Molecular Modules

Membrane-association determinants of the ω-amino acid monooxygenase PvdA, a pyoverdine biosynthetic enzyme from Pseudomonas aeruginosa

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