Xanthomonas campestris pv. vesicatoria Secretes Proteases and Xylanases via the Xps Type II Secretion System and Outer Membrane Vesicles

Solé, Magali; Scheibner, Felix; Hoffmeister, Anne-Katrin; Hartmann, N; Hause, Gerd; Rother, Annekatrin; Jordan, Michael; Lautier, Martine; Arlat, Matthieu; Büttner, Daniela

doi:10.1128/jb.00322-15

Cited by 82 publications

(82 citation statements)

References 68 publications

(98 reference statements)

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“…On evolutionary time scales, public goods provided by OMVs may select for cooperative behavior, driving the establishment of mutualisms. Finally, we note that the preponderance of research has examined OMVs in the context of animal symbioses; to date, only a few studies have focused on OMV-plant interactions (23,(99)(100)(101)(102)(103). Extending the role of OMVs in the biology of other multicellular lineages is a budding area for future investigation.…”

Section: Discussionmentioning

confidence: 99%

Spheres of Hope, Packets of Doom: the Good and Bad of Outer Membrane Vesicles in Interspecies and Ecological Dynamics

Lynch

Alegado

2017

J Bacteriol

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Outer membrane vesicles (OMVs) are proteoliposome nanoparticles ubiquitously produced by Gram-negative bacteria. Typically bearing a composition similar to those of the outer membrane and periplasm of the cells from which they are derived, OMVs package an array of proteins, lipids, and nucleic acids. Once considered inconsequential by-products of bacterial growth, OMVs have since been demonstrated to mediate cellular stress relief, promote horizontal gene transfer and antimicrobial activity, and elicit metazoan inflammation. Recently, OMVs have gained appreciation as critical moderators of interorganismal dynamics. In this review, we focus on recent progress toward understanding the functions of OMVs with regard to symbiosis and ecological contexts, and we propose potential avenues for future OMV studies.

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Section: Discussionmentioning

confidence: 99%

Spheres of Hope, Packets of Doom: the Good and Bad of Outer Membrane Vesicles in Interspecies and Ecological Dynamics

Lynch

Alegado

2017

J Bacteriol

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“…Horseradish peroxidase-labeled anti-rabbit or anti-mouse antibodies (GE Healthcare) were used as secondary antibodies. For the analysis of protein secretion via outer membrane vesicles, bacteria were incubated on a rotary shaker overnight at 30°C and supernatants were incubated with proteinase K as described previously (44). Experiments were performed three times with similar results.…”

Section: Methodsmentioning

confidence: 99%

“…For the analysis of OMVmediated protein secretion, strain 85-10hpaHoof containing HpaH-c-Myc was cultivated in nutrient-yeast-glycerol (NYG) medium, and bacterial culture supernatants were incubated in the absence or presence of proteinase K. OMV cargo proteins are protected from degradation by proteinase K, as is observed for the T2S substrate XCV4360 -c-Myc (Fig. 5B) (44). However, this was not the case for HpaH-c-Myc, suggesting that HpaH is secreted independently of OMVs by an alternative transport pathway.…”

Section: (B)mentioning

confidence: 99%

“…OMVs from X. campestris pv. vesicatoria were previously shown to contain predicted virulence factors, including T2S substrates, suggesting that periplasmic proteins can be targeted to different transport routes (44). For the analysis of OMVmediated protein secretion, strain 85-10hpaHoof containing HpaH-c-Myc was cultivated in nutrient-yeast-glycerol (NYG) medium, and bacterial culture supernatants were incubated in the absence or presence of proteinase K. OMV cargo proteins are protected from degradation by proteinase K, as is observed for the T2S substrate XCV4360 -c-Myc (Fig.…”

Section: (B)mentioning

confidence: 99%

“…T2S systems are Sec-dependent protein transport systems which often secrete virulence factors across the outer bacterial membrane (41,42). We previously reported that the Xps T2S system contributes to virulence and secretes degradative enzymes (43,44). For secretion assays with HpaH-c-Myc, we used the wild-type strain 85-10 and the T2S-deficient strain 85-10ΔEEE, which lacks three putative ATPase-encoding genes of both T2S systems (43).…”

Section: (B)mentioning

confidence: 99%

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The Predicted Lytic Transglycosylase HpaH from Xanthomonas campestris pv. vesicatoria Associates with the Type III Secretion System and Promotes Effector Protein Translocation

et al. 2017

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The pathogenicity of the Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion (T3S) system, which spans both bacterial membranes and translocates effector proteins into plant cells. The assembly of the T3S system presumably involves the predicted lytic transglycosylase (LT) HpaH, which is encoded adjacent to the T3S gene cluster. Bacterial LTs degrade peptidoglycan and often promote the formation of membrane-spanning macromolecular protein complexes. In the present study, we show that HpaH localizes to the bacterial periplasm and binds to peptidoglycan as well as to components of the T3S system, including the predicted periplasmic inner rod proteins HrpB1 and HrpB2 as well as the pilus protein HrpE. In vivo translocation assays revealed that HpaH promotes the translocation of various effector proteins and of early substrates of the T3S system, suggesting a general contribution of HpaH to type III-dependent protein export. Mutant studies and the analysis of reporter fusions showed that the N-terminal region of HpaH contributes to protein function and is proteolytically cleaved. The N-terminally truncated HpaH cleavage product is secreted into the extracellular milieu by a yet-unknown transport pathway, which is independent of the T3S system. KEYWORDS Xanthomonas, peptidoglycan, Slt70, type III secretion, lytic transglycosylase, effector proteins T he pathogenicity of many Gram-negative plant-and animal-pathogenic bacteria depends on a type III secretion (T3S) system, which translocates bacterial effector proteins into eukaryotic cells (1). Type III effector proteins manipulate host cellular pathways to the benefit of the pathogen and thus promote bacterial proliferation (2). T3S systems are highly complex nanomachines that span both bacterial membranes and are associated with a pilus-like appendage, which serves as a transport channel for secreted proteins (3). Protein translocation into eukaryotic cells depends on the channel-like T3S translocon, which inserts into the host plasma membrane (3, 4).T3S systems usually consist of 20 to 25 proteins, nine of which are conserved in plant-and animal-pathogenic bacteria and constitute the core elements of the secretion apparatus. These include ring structures in the bacterial inner membrane (IM) and outer membrane (OM), which are associated with a predicted periplasmic inner rod (3). The IM ring interacts with components of the export apparatus, which is assembled by members of the conserved Sct (secretion and cellular translocation) R, S, T, U, and V families of transmembrane proteins (3). The letters refer to the Ysc proteins of the T3S system from Yersinia spp. (5, 6). Components of the export apparatus insert into the IM and interact with cytoplasmic parts of the T3S system, including the predicted cytoplasmic (C) ring and the ATPase complex, which is presumably involved in T3S substrate

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Bacterial outer membrane vesicles induce a transcriptional shift in arabidopsis towards immune system activation leading to suppression of pathogen growth in planta

Chalupowicz

Mordukhovich

Assoline

et al. 2023

J of Extracellular Vesicle

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Gram-negative bacteria form spherical blebs on their cell periphery, which later dissociate from the bacterial cell wall to form extracellular vesicles. These nano scale structures, known as outer membrane vesicles (OMVs), have been shown to promote infection and disease and can induce typical immune outputs in both mammal and plant hosts. To better understand the broad transcriptional change plants undergo following exposure to OMVs, we treated Arabidopsis thaliana (Arabidopsis) seedlings with OMVs purified from the Gram-negative plant pathogenic bacterium Xanthomonas campestris pv. campestris and performed RNA-seq analysis on OMVand mock-treated plants at 2, 6 and 24 h post challenge. The most pronounced transcriptional shift occurred at the first two time points tested, as reflected by the number of differentially expressed genes and the average fold change. OMVs induce a major transcriptional shift towards immune system activation, upregulating a multitude of immune-related pathways including a variety of immune receptors. Comparing the response of Arabidopsis to OMVs and to purified elicitors, revealed that OMVs induce a similar suite of genes and pathways as single elicitors, however, pathways activated by OMVs and not by other elicitors were detected. Pretreating Arabidopsis plants with OMVs and subsequently infecting with a bacterial pathogen led to a significant reduction in pathogen growth. Mutations in the plant elongation factor receptor (EFR), flagellin receptor (FLS2), or the brassinosteroid-insensitive 1associated kinase (BAK1) co-receptor, did not significantly affect the immune priming effect of OMVs. All together these results show that OMVs induce a broad transcriptional shift in Arabidopsis leading to upregulation of multiple immune pathways, and that this transcriptional change may facilitate resistance to bacterial infection.

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Xanthomonas campestris pv. vesicatoria Secretes Proteases and Xylanases via the Xps Type II Secretion System and Outer Membrane Vesicles

Cited by 82 publications

References 68 publications

Spheres of Hope, Packets of Doom: the Good and Bad of Outer Membrane Vesicles in Interspecies and Ecological Dynamics

Spheres of Hope, Packets of Doom: the Good and Bad of Outer Membrane Vesicles in Interspecies and Ecological Dynamics

The Predicted Lytic Transglycosylase HpaH from Xanthomonas campestris pv. vesicatoria Associates with the Type III Secretion System and Promotes Effector Protein Translocation

Bacterial outer membrane vesicles induce a transcriptional shift in arabidopsis towards immune system activation leading to suppression of pathogen growth in planta

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