A distinct inhibitory mechanism of the V-ATPase by Vibrio VopQ revealed by cryo-EM

Peng, Wei; Casey, Amanda K.; Fernandez, Jessie; Carpinone, Emily M.; Servage, Kelly A.; Chen, James Z.; Li, Yang; Tomchick, Diana R.; Starai, Vincent J.; Orth, Kim

doi:10.1038/s41594-020-0429-1

Cited by 10 publications

(20 citation statements)

References 65 publications

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“…A Vibrio parahaemolyticus type 3 secretion system effector, VopQ, was previously shown to bind dissociated Vo subcomplex and predicted to bind the holoenzyme of V1Vo in state 2 (ref. 17 ).…”

Section: V1vo Holoenzyme In Complex With a Bacterial Effectormentioning

confidence: 99%

“…However, the density maps for VopQ were not well resolved, indicating that VopQ may partially clashed with subunits in V1Vo and the relatively dynamic Vo regions may also contribute to this clash. A VopQ molecule corresponding to VopQ-2B in the reported Vo-VopQ structures 17 was docked into the cryo-EM density map ( Fig. 2a).…”

Section: V1vo Holoenzyme In Complex With a Bacterial Effectormentioning

confidence: 99%

“…The structures of yeast V1Vo (PDB code: 3J9U) 7 and Vo-VopQ (PDB code: 6PE5) 17 were used as initial models for model building of V1Vo-VopQ (state2). The model was manually adjusted in COOT 38 .…”

Section: Model Building and Structure Refinementmentioning

confidence: 99%

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Structures of an intact yeast V-ATPase alone and in complex with bacterial effector VopQ

Orth

Peng

Casey

et al. 2020

Preprint

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Vacuolar-type ATPase (V-ATPase) is a rotary protein pump involved in proton translocation across various cellular membranes using the energy of ATP hydrolysis. Despite previous studies on bacterial and eukaryotic V-ATPases, information on the intact structure of a eukaryotic V-ATPase is missing. Here we report cryo-EM structures of the intact yeast V-ATPase and this complex bound to a bacterial effector. We reveal the interaction of the elusive regulatory subunit H with its neighboring subunits. Insight for the catalytic mechanism is gained by determining conformations of the catalytic subunits either empty or bound with nucleotides.

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Section: V1vo Holoenzyme In Complex With a Bacterial Effectormentioning

confidence: 99%

Section: V1vo Holoenzyme In Complex With a Bacterial Effectormentioning

confidence: 99%

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Structures of an intact yeast V-ATPase alone and in complex with bacterial effector VopQ

Orth

Peng

Casey

et al. 2020

Preprint

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“…Recently, cryo-EM studies performed in our lab revealed extensive interactions between VopQ and the c subunit of V o V-ATPase that stabilize the insertion of VopQ in the membrane alongside the V-ATPase (16). The interactions of VopQ with the c-ring of V o is predicted to form an unconventional membrane pore through the juxtaposition of charged resides of VopQ against the hydrophobic lipid environment (16). This disruption is predicted to lead to the deacidification of the lysosomal membrane.…”

Section: Introductionmentioning

confidence: 99%

“…Remarkably, a fully functioning or assembled V-ATPase at the vacuole is not necessary to induce VopQ toxicity in yeast. We found that VopQ can interact with an assembly intermediate of the V-ATPase (V o c-ring) in the ER resulting in cell death (16). As VopQ forms a pore in target membranes, the ER membrane is compromised, and this could lead to the induction of host cell signaling events including the unfolded protein response (UPR).…”

Section: Introductionmentioning

confidence: 99%

Manipulation of IRE1-dependent MAPK signaling by a Vibrio agonist-antagonist effector pair

Nisco

Casey

Kanchwala

et al. 2020

Preprint

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Diverse bacterial pathogens employ effector delivery systems to disrupt vital cellular processes in the host (1). The type III secretion system 1 of the marine pathogen, Vibrio parahaemolyticus, utilizes the sequential action of four effectors to induce a rapid, pro-inflammatory cell death uniquely characterized by a pro-survival host transcriptional response (2, 3). Herein, we show that this pro-survival response is caused by the action of the channel-forming effector VopQ that targets the host V-ATPase resulting in lysosomal deacidification and inhibition of lysosome-autophagosome fusion. Recent structural studies have shown how VopQ interacts with the V-ATPase and, while in the ER, a V-ATPase assembly intermediate can interact with VopQ causing a disruption in membrane integrity. Additionally, we observe that VopQ-mediated disruption of the V-ATPase activates the IRE1 branch of the unfolded protein response (UPR) resulting in an IRE1-dependent activation of ERK1/2 MAPK signaling. We also find that this early VopQ-dependent induction of ERK1/2 phosphorylation is terminated by the VopS-mediated inhibitory AMPylation of Rho GTPase signaling. Since VopS dampens VopQ-induced IRE1-dependent ERK1/2 activation, we propose that IRE1 activates ERK1/2 phosphorylation at or above the level of Rho GTPases. This study illustrates how temporally induced effectors can work as in tandem as agonist/antagonist to manipulate host signaling and reveal new connections between V-ATPase function, UPR and MAPK signaling.ImportanceVibrio parahaemolyticus (V. para) is a seafood-borne pathogen that encodes two Type 3 Secretion Systems (T3SS). The first system T3SS1 is thought to be maintained in all strains of V. para to to maintain survival in the environment, whereas the second sytem T3SS2 is linked to clinical isolates and disease in humans. Herein, we find that first system targets evolutionarily conserved signaling systems to manipulate host cells, eventually causing a rapid, orchestrated cells death within three hours. We have found that the T3SS1 injects virulence factors that temporally manipulate host signaling. Within the first hour of infection, the effector VopQ acts first by activating host surval signals while diminishing the host cell apoptotic machinery. Less than an hour later, another effector VopS reverses activation and inhibition of these signaling systems ultimately leading to death of the host cell. This work provides example of how pathogens have evolved to manipulate the interplay between T3SS effectors to regulate host signaling pathways.

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Unravelling bacterial virulence factors in yeast: From identification to the elucidation of their mechanisms of action

Wong,

Ong

2024

Arch Microbiol

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A distinct inhibitory mechanism of the V-ATPase by Vibrio VopQ revealed by cryo-EM

Cited by 10 publications

References 65 publications

Structures of an intact yeast V-ATPase alone and in complex with bacterial effector VopQ

Structures of an intact yeast V-ATPase alone and in complex with bacterial effector VopQ

Manipulation of IRE1-dependent MAPK signaling by a Vibrio agonist-antagonist effector pair

Unravelling bacterial virulence factors in yeast: From identification to the elucidation of their mechanisms of action

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