Structure of a bacterial type IV secretion core complex at subnanometre resolution

Rivera-Calzada, Ángel; Fronzes, Rémi; Savva, Christos G.; Chandran, Vidya; Lian, Pei W; Laeremans, Toon; Pardon, Els; Steyaert, Jan; Remaut, Han; Waksman, Gabriel; Orlova, Elena V.

doi:10.1038/emboj.2013.58

Cited by 87 publications

(113 citation statements)

References 35 publications

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“…In this system, T4S system components are termed VirB1‐VirB11 and VirD4 (Fronzes et al , 2009a). The central channel of the T4S system, called the “core” complex, is formed from 14 copies of the proteins VirB7, VirB9 and VirB10, and spans both membranes (Fronzes et al , 2009b; Rivera‐Calzada et al , 2013). The overall structure of the core complex appears as a two‐layered ring with 14‐fold symmetry.…”

Section: Introductionmentioning

confidence: 99%

Structure of a VirD4 coupling protein bound to a VirB type IV secretion machinery

et al. 2017

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Type IV secretion (T4S) systems are versatile bacterial secretion systems mediating transport of protein and/or DNA. T4S systems are generally composed of 11 VirB proteins and 1 VirD protein (VirD4). The VirB1‐11 proteins assemble to form a secretion machinery and a pilus while the VirD4 protein is responsible for substrate recruitment. The structure of VirD4 in isolation is known; however, its structure bound to the VirB1‐11 apparatus has not been determined. Here, we purify a T4S system with VirD4 bound, define the biochemical requirements for complex formation and describe the protein–protein interaction network in which VirD4 is involved. We also solve the structure of this complex by negative stain electron microscopy, demonstrating that two copies of VirD4 dimers locate on both sides of the apparatus, in between the VirB4 ATPases. Given the central role of VirD4 in type IV secretion, our study provides mechanistic insights on a process that mediates the dangerous spread of antibiotic resistance genes among bacterial populations.

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Section: Introductionmentioning

confidence: 99%

Structure of a VirD4 coupling protein bound to a VirB type IV secretion machinery

et al. 2017

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“…Recently, an almost complete T4SS, encoded by the R388 plasmid, was purified and visualised using electron microscopy 106 (FIG. 3c) The OMC core complex is formed of two layers, one of which (termed "the Olayer") inserts in the OM 107,108 . It is not known how the T4SS apparatus assembles a conjugative pilus.…”

Section: Structure Of the Helical Filamentmentioning

confidence: 99%

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

2017

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Pili are critical virulence factors of many Gram-negative pathogens. These surface structures provide bacteria with a link to their outside environments, allowing bacteria to interact with their hosts, other surfaces or with each other. They can even provide a conduit for the exchange of information. The surface chemistries and other biophysical properties of pili are uniquely adapted to the environmental challenges faced by bacteria. The assembly of these surface structures presents a molecular engineering challenge, which bacteria have solved in a variety of unique ways. In this review, we examine recent advances in our structural understanding of various Gram-negative pilus systems and discuss their functional implications.

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“…The translocation of a conjugative plasmid relies on the coupling protein, the relaxosome complex, and a type IV secretion system (T4SS), known as the mating-pair formation (Mpf) apparatus (1). For conjugation systems in Gram-negative bacteria, structural resolution of the outer membrane core complex and the inner membrane complex of the T4SS, as well as information about the pathway of DNA translocation across the T4SS in the donor cell, has been elucidated (3)(4)(5)(6)(7)(8)(9)(10)(11). Less is known about conjugation systems in Gram-positive bacteria, with our current understanding based primarily on comparisons to those in Gramnegative bacteria and with the systems in Gram-positive bacteria proposed to involve a "minimized" T4SS (3,(12)(13)(14)(15)(16)(17)(18).…”

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Two Novel Membrane Proteins, TcpD and TcpE, Are Essential for Conjugative Transfer of pCW3 in Clostridium perfringens

et al. 2015

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The anaerobic pathogen Clostridium perfringens encodes either toxin genes or antibiotic resistance determinants on a unique family of conjugative plasmids that have a novel conjugation region, the tcp locus. Studies of the paradigm conjugative plasmid from C. perfringens, the 47-kb tetracycline resistance plasmid pCW3, have identified several tcp-encoded proteins that are involved in conjugative transfer and form part of the transfer apparatus. In this study, the role of the conserved hypothetical proteins TcpD, TcpE, and TcpJ was examined. Mutation and complementation analyses showed that TcpD and TcpE were essential for the conjugative transfer of pCW3, whereas TcpJ was not required. To analyze the TcpD and TcpE proteins in C. perfringens, functional hemagglutinin (HA)-tagged derivatives were constructed. Western blots showed that TcpD and TcpE localized to the cell envelope fraction independently of the presence of other pCW3-encoded proteins. Finally, examination of the subcellular localization of TcpD and TcpE by immunofluorescence showed that these proteins were concentrated at both poles of C. perfringens donor cells, where they are postulated to form essential components of the multiprotein complex that comprises the transfer apparatus. Conjugative transfer of plasmid DNA is an important process that contributes to bacterial genome plasticity and adaption, particularly the spread of antibiotic resistance and virulence genes (1, 2). The translocation of a conjugative plasmid relies on the coupling protein, the relaxosome complex, and a type IV secretion system (T4SS), known as the mating-pair formation (Mpf) apparatus (1). For conjugation systems in Gram-negative bacteria, structural resolution of the outer membrane core complex and the inner membrane complex of the T4SS, as well as information about the pathway of DNA translocation across the T4SS in the donor cell, has been elucidated (3-11). Less is known about conjugation systems in Gram-positive bacteria, with our current understanding based primarily on comparisons to those in Gramnegative bacteria and with the systems in Gram-positive bacteria proposed to involve a "minimized" T4SS (3, 12-18).The Gram-positive, anaerobic pathogen Clostridium perfringens carries many disease-mediating toxin genes and antibiotic resistance genes on a unique family of large plasmids that have large regions of sequence identity (19)(20)(21)(22). Most of these plasmids carry the novel tcp conjugation locus (20, 23). The tcp locus has been identified on all known conjugative plasmids in C. perfringens, and proteins encoded by this locus have been shown to be required for conjugative transfer of the paradigm C. perfringens conjugative plasmid, pCW3 (20,(23)(24)(25)(26). The presence of the tcp genes on so many of these toxin plasmids suggests that the transfer of toxin genes by a shared conjugation mechanism in C. perfringens is important in disease dissemination. To this effect, several of these toxin plasmids have been shown experimentally to be conjugative (19,27,28).M...

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Structure of a bacterial type IV secretion core complex at subnanometre resolution

Cited by 87 publications

References 35 publications

Structure of a VirD4 coupling protein bound to a VirB type IV secretion machinery

Structure of a VirD4 coupling protein bound to a VirB type IV secretion machinery

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

Two Novel Membrane Proteins, TcpD and TcpE, Are Essential for Conjugative Transfer of pCW3 in Clostridium perfringens

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