New Insights into the Assembly of Bacterial Secretins

Meeren, Ruben Van der; Wen, Yurong; Gelder, Patrick Van; Tommassen, Jan; Devreese, Bart; Savvides, Savvas N.

doi:10.1074/jbc.m112.432096

Cited by 28 publications

(43 citation statements)

References 40 publications

(45 reference statements)

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“…Also several crystal structures of periplasmic GspD domains have been reported. These include the structures of the N0-N1-N2 domains of ETEC GspD in complex with a nanobody (Korotkov et al, 2009), the N0-N1 domains of ETEC GspD in complex with the HR-domain of GspC (Korotkov et al, 2011b), and the N0-N1-N2 domains of Pseudomonas aeruginosa GspD XcpQ (Van der Meeren et al, 2013). These structures all share a similar compact N0-N1 module.…”

Section: Introductionmentioning

confidence: 99%

“…A combination of the crystal structures of individual secretin domains with the cryo-electron microscopy reconstruction resulted in dodecameric models with C 12 symmetry for the organization of the periplasmic domains of GspD (Korotkov et al, 2011b; Reichow et al, 2010). In contrast, a hexameric assembly of dimers has been suggested based on the basis of cysteine scanning mutagenesis and disulfide mapping analysis of Dickeya dadantii (previously Erwinia chrysanthemi ) GspD OutD (Wang et al, 2012), and also on the basis of dimers occurring in the crystal structure of the N0-N1-N2 domains of P. aeruginosa GspD XcpQ (Van der Meeren et al, 2013). So far, however, high-resolution experimental information on the multimeric structure of a secretin is still missing.…”

Section: Introductionmentioning

confidence: 99%

“…Based on this helical organization, a model of a N0 ring with C 12 point group symmetry is obtained whose dimensions agree with the cryo-electron microscopy reconstruction (Reichow et al, 2010). The dodecameric N0-GspD ring is analyzed in the light of numerous recent reports regarding the structure and function of the N0 domain and the interactions of the N0 domain of GspD with the HR-domain of the inner membrane T2SS protein GspC (Douzi et al, 2011; Gu et al, 2012; Korotkov et al, 2009; Korotkov et al, 2011b; Login et al, 2010; Van der Meeren et al, 2013; Wang et al, 2012). Our analysis suggests that the N0 domain of the T2SS secretin has an intrinsic tendency to form a cylindrical dodecamer, and that, consequently, T2SS and related secretins are multimers with cylindrical 12-fold symmetry.…”

Section: Introductionmentioning

confidence: 99%

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A dodecameric ring-like structure of the N0 domain of the type II secretin from enterotoxigenic Escherichia coli

Korotkov

Hól

2013

Journal of Structural Biology

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In many bacteria, secretins from the type II secretion system (T2SS) function as outer membrane gated channels that enable passage of folded proteins from the periplasm into the extracellular milieu. Cryo-electron microscopy of the T2SS secretin GspD revealed previously the dodecameric cylindrical architecture of secretins, and crystal structures of periplasmic secretin domains showed a modular domain organization. However, no high-resolution experimental data has as yet been provided about how the entire T2SS secretin or its domains are organized in a cylindrical fashion. Here we present a crystal structure of the N0 domain of the T2SS secretin GspD from enterotoxigenic Escherichia coli containing a helix with 12 subunits per turn. The helix has an outer diameter of ~125 Å and a pitch of only 24 Å which suggests a model of a cylindrical dodecameric N0 ring whose dimensions correspond with the cryo-electron microscopy map of Vibrio cholerae GspD. The N0 domain is known to interact with the HR domain of the inner membrane T2SS protein GspC. When the new N0 ring model is combined with the known N0• HR crystal structure, a dodecameric double-ring of twelve N0-HR heterodimers is obtained. In contrast, the previously observed compact N0-N1 GspD module is not compatible with the N0 ring. Interestingly, a N0-N1 T3SS homolog is compatible with forming a N0-N1 dodecameric ring, due to a different N0-vs-N1 orientation. This suggests that the dodecameric N0 ring is an important feature of T2SS secretins with periplasmic domains undergoing considerable motions during exoprotein translocation.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A dodecameric ring-like structure of the N0 domain of the type II secretin from enterotoxigenic Escherichia coli

Korotkov

Hól

2013

Journal of Structural Biology

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“…To our knowledge, multimeric forms of PilP and PilQ of T4PS have not been reported previously. The periplasmic domain of the XcpD secretin of the P. aeruginosa T2SS was reported to form a dimer (71).…”

Section: Stability Of T4ps Proteins In the Absence Of Each Individualmentioning

confidence: 99%

Outside-In Assembly Pathway of the Type IV Pilus System in Myxococcus xanthus

2014

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“…The N domain is located in the periplasm and interacts with inner membrane components of the secretion machinery (6)(7)(8)(9). The atomic resolution structures of part of the N domains of several secretins have been solved by X-ray crystallography (for the T3SS, EscC [10], and for the T2SS, GspD [11] and XcpQ [12]) and nuclear magnetic resonance (NMR) spectroscopy (for T4P, EscC [13]). Some secretins, including PulD, possess a C-terminal extension (S domain) (Fig.…”

mentioning

confidence: 99%

Bacterial Secretins Form Constitutively Open Pores Akin to General Porins

Disconzi¹,

Guilvout²,

Chami³

et al. 2013

Journal of Bacteriology

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e Proteins called secretins form large multimeric complexes that are essential for macromolecular transit across the outer membrane of Gram-negative bacteria. Evidence suggests that the channels formed by some secretin complexes are not tightly closed, but their permeability properties have not been well characterized. Here, we used cell-free synthesis coupled with spontaneous insertion into liposomes to investigate the permeability of the secretin PulD. Leakage assays using preloaded liposomes indicated that PulD allows the efflux of small fluorescent molecules with a permeation cutoff similar to that of general porins. Other secretins were also found to form similar pores. To define the polypeptide region involved in determining the pore size, we analyzed a collection of PulD variants and studied the roles of gates 1 and 2, which were previously reported to affect the pore size of filamentous phage f1 secretin pIV, in assembly and pore formation. Liposome leakage and a novel in vivo assay showed that replacement of the conserved proline residue at position 443 in PulD by leucine increased the apparent size of the pore. The in vitro approach described here could be used to study the pore properties of membrane proteins whose production in vivo is toxic.

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New Insights into the Assembly of Bacterial Secretins

Cited by 28 publications

References 40 publications

A dodecameric ring-like structure of the N0 domain of the type II secretin from enterotoxigenic Escherichia coli

A dodecameric ring-like structure of the N0 domain of the type II secretin from enterotoxigenic Escherichia coli

Outside-In Assembly Pathway of the Type IV Pilus System in Myxococcus xanthus

Bacterial Secretins Form Constitutively Open Pores Akin to General Porins

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