<i>Photorhabdus luminescens</i>
            Toxins ADP-Ribosylate Actin and RhoA to Force Actin Clustering

Lang, Alexander E.; Schmidt, Gudula; Schlösser, Andreas; Hey, T D; Larrinua, Ignacio M.; Sheets, Joel J.; Mannherz, Hans Georg; Aktories, Klaus

doi:10.1126/science.1184557

Cited by 217 publications

(300 citation statements)

References 23 publications

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“…This proposal is supported by small-angle X-ray scattering data, which are consistent with a hollow spheroid for the B/C NTR complex, but with a solid spheroid for proteases, can be safely expressed without causing damage to the producing cell. A plausible explanation supported by this structure would be that the toxic payload remains sequestered until exposure to a change in pH triggers its release 5 (Fig. 2a).…”

mentioning

confidence: 92%

The BC component of ABC toxins is an RHS-repeat-containing protein encapsulation device

Busby

Panjikar

Landsberg

et al. 2013

Nature

140

198

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The ABC toxin complexes (Tc) produced by certain bacteria are of interest due to their potent insecticidal activity 1,2 and potential role in human disease 3 . These complexes comprise at least three proteins (A, B and C), which must assemble to be fully toxic 4 . The carboxy-terminal region of C is the main cytotoxic component 5 , and is poorly conserved between different Tcs. A general model of action has been proposed, in which the Tc binds to the cell surface via the A protein, is endocytosed, and subsequently forms a pH-triggered channel, allowing the translocation of C into the cytoplasm, where it can cause cytoskeletal disruption in both insect and mammalian cells 5 . Tc complexes have been visualised using single particle electron microscopy 6,7 , but no high-resolution structures of the components are available, and the role of the B protein in the mechanism of toxicity remains unknown. Here we report the three-dimensional structure of the complex formed between the B and C proteins, determined to 2.5 Å by X-ray crystallography. These proteins assemble to form an unprecedented, large hollow structure that encapsulates and sequesters the cytotoxic, carboxy-terminal region of the C protein like the shell of an egg. The shell is decorated on one end with a -propeller domain, which mediates attachment of the B/C heterodimer to the A protein in the native complex. The structure reveals how C auto-proteolyses when folded in complex with B. The C protein is the first example of a structure that contains RHS (rearrangement hot spot) repeats 8 , and illustrates a striking structural architecture that is likely conserved across both this widely distributed bacterial protein family and the related eukaryotic YD-repeatcontaining protein family, which includes the teneurins 9 . The structure provides the first clues about the function of these protein repeat families, and suggests a generic mechanism for protein encapsulation and delivery.ABC toxins were first identified, and have been best characterised, in the bacterium Photorhabdus luminescens 1 . However, the entomopathogenic bacterium Yersinia entomophaga contains a related Tc locus that includes an A component encoded by two ORFs (yenA1 and yenA2), a single B gene (yenB) and two C genes (yenC1 and yenC2), 10 the products of which associate independently with the A and B proteins, giving rise to two Tcs from one genetic locus. The C proteins of this and other Tcs are similar to the "polymorphic toxins" described by Zhang et al. 11 as they have a conserved RHS-repeat-containing amino-terminal region and a variable carboxyterminal region 10 . The carboxy-terminal regions (CTRs) of the Y. entomophaga C proteins are predicted to have different toxic activities: the C1 CTR is homologous to cytotoxic necrotising factor 1 (CNF1) from Escherichia coli 12 , whereas the C2 CTR is homologous to the deaminase YwqJ from Bacillus subtilis 13 . As in related complexes 3 , when the B subunit is co-expressed with either of the C subunits, C is cleaved at the boundary between th...

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mentioning

confidence: 92%

The BC component of ABC toxins is an RHS-repeat-containing protein encapsulation device

Busby

Panjikar

Landsberg

et al. 2013

Nature

140

198

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“…Further evidence for this is provided by the observation that the TcA components of an insecticidal X. nematophila Tc bind to solubilized insect midgut brush border membranes (14,15). By comparison, TcC-like proteins are thought to represent the main toxin components of the Tc family (7,16). Two P. luminescens TcC-like toxins, TccC3 and TccC5, trigger ADP ribosylation and RhoA GTPase activation in Galleria melonella haemocytes and cultured HeLa cells, inducing redistribution and clustering of actin, inhibiting phagocytosis and leading to cell death (16).…”

mentioning

confidence: 97%

3D structure of the Yersinia entomophaga toxin complex and implications for insecticidal activity

Landsberg

Jones

Rothnagel

et al. 2011

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

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Toxin complex (Tc) proteins are a class of bacterial protein toxins that form large, multisubunit complexes. Comprising TcA, B, and C components, they are of great interest because many exhibit potent insecticidal activity. Here we report the structure of a novel Tc, Yen-Tc, isolated from the bacterium Yersinia entomophaga MH96, which differs from the majority of bacterially derived Tcs in that it exhibits oral activity toward a broad range of insect pests, including the diamondback moth ( Plutella xylostella ). We have determined the structure of the Yen-Tc using single particle electron microscopy and studied its mechanism of toxicity by comparative analyses of two variants of the complex exhibiting different toxicity profiles. We show that the A subunits form the basis of a fivefold symmetric assembly that differs substantially in structure and subunit arrangement from its most well characterized homologue, the Xenorhabdus nematophila toxin XptA1. Histopathological and quantitative dose response analyses identify the B and C subunits, which map to a single, surface-accessible region of the structure, as the sole determinants of toxicity. Finally, we show that the assembled Yen-Tc has endochitinase activity and attribute this to putative chitinase subunits that decorate the surface of the TcA scaffold, an observation that may explain the oral toxicity associated with the complex.

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“…the cholera toxin modifies GasR187 28 -to alter the behavior of host proteins. Indeed, bacterial toxin ARTs have evolved a wide range of substrates, such as the Bordetella pertussis toxin that modifies cysteine, the Clostridium botulinum C3 toxin that modifies asparagine, the Photorhabdus luminescens toxin that modifies glutamine 29 and the Corynebacterium diphtheriae toxin that modifies diphthamide. 30 In contrast, the lepidopteran ARTs, such as pierisin, modify the N2 amino group of guanine in DNA to induce apoptosis.…”

Section: Introductionmentioning

confidence: 99%

Identification of novel components of NAD-utilizing metabolic pathways and prediction of their biochemical functions

Souza

Aravind

2012

Mol. BioSyst.

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Nicotinamide adenine dinucleotide (NAD) is a ubiquitous cofactor participating in numerous redox reactions. It is also a substrate for regulatory modifications of proteins and nucleic acids via the addition of ADP-ribose moieties or removal of acyl groups by transfer to ADP-ribose. In this study, we use in-depth sequence, structure and genomic context analysis to uncover new enzymes and substrate-binding proteins in NAD-utilizing metabolic and macromolecular modification systems. We predict that Escherichia coli YbiA and related families of domains from diverse bacteria, eukaryotes, large DNA viruses and single strand RNA viruses are previously unrecognized components of NAD-utilizing pathways that probably operate on ADP-ribose derivatives. Using contextual analysis we show that some of these proteins potentially act in RNA repair, where NAD is used to remove 2 0 -3 0 cyclic phosphodiester linkages. Likewise, we predict that another family of YbiA-related enzymes is likely to comprise a novel NAD-dependent ADP-ribosylation system for proteins, in conjunction with a previously unrecognized ADP-ribosyltransferase. A similar ADP-ribosyltransferase is also coupled with MACRO or ADP-ribosylglycohydrolase domain proteins in other related systems, suggesting that all these novel systems are likely to comprise pairs of ADP-ribosylation and ribosylglycohydrolase enzymes analogous to the DraG-DraT system, and a novel group of bacterial polymorphic toxins. We present evidence that some of these coupled ADP-ribosyltransferases/ribosylglycohydrolases are likely to regulate certain restriction modification enzymes in bacteria. The ADP-ribosyltransferases found in these, the bacterial polymorphic toxin and host-directed toxin systems of bacteria such as Waddlia also throw light on the evolution of this fold and the origin of eukaryotic polyADP-ribosyltransferases and NEURL4-like ARTs, which might be involved in centrosomal assembly. We also infer a novel biosynthetic pathway that might be involved in the synthesis of a nicotinate-derived compound in conjunction with an asparagine synthetase and AMPylating peptide ligase. We use the data derived from this analysis to understand the origin and early evolutionary trajectories of key NAD-utilizing enzymes and present targets for future biochemical investigations.

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Photorhabdus luminescens Toxins ADP-Ribosylate Actin and RhoA to Force Actin Clustering

Cited by 217 publications

References 23 publications

The BC component of ABC toxins is an RHS-repeat-containing protein encapsulation device

The BC component of ABC toxins is an RHS-repeat-containing protein encapsulation device

3D structure of the Yersinia entomophaga toxin complex and implications for insecticidal activity

Identification of novel components of NAD-utilizing metabolic pathways and prediction of their biochemical functions

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