@article{c19c7ba43e024515918891ce33f7f9ed,
title = "A distinct inhibitory mechanism of the V-ATPase by Vibrio VopQ revealed by cryo-EM",
abstract = "The Vibrioparahaemolyticus T3SS effector VopQ targets host-cell V-ATPase, resulting in blockage of autophagic flux and neutralization of acidic compartments. Here, we report the cryo-EM structure of VopQ bound to the Vo subcomplex of the V-ATPase. VopQ inserts into membranes and forms an unconventional pore while binding directly to subunit c of the V-ATPase membrane-embedded subcomplex Vo. We show that VopQ arrests yeast growth in vivo by targeting the immature Vo subcomplex in the endoplasmic reticulum (ER), thus providing insight into the observation that VopQ kills cells in the absence of a functional V-ATPase. VopQ is a bacterial effector that has been discovered to inhibit a host-membrane megadalton complex by coincidentally binding its target, inserting into a membrane and disrupting membrane potential. Collectively, our results reveal a mechanism by which bacterial effectors modulate host cell biology and provide an invaluable tool for future studies on V-ATPase-mediated membrane fusion and autophagy.",
author = "Wei Peng and Casey, {Amanda K.} and Jessie Fernandez and Carpinone, {Emily M.} and Servage, {Kelly A.} and Zhe Chen and Yang Li and Tomchick, {Diana R.} and Starai, {Vincent J.} and Kim Orth",
note = "Funding Information: We thank V. Tagliabracci, A. Sreelatha, X. Bai, X. Li and members of the Orth laboratory for discussions and editing. We thank X. Bai for technical assistance with cryo-EM image processing. We thank the Friedman laboratory for assistance with yeast cell disruption. We thank the Electron Microscopy Core Facility for assistance with negative staining grid preparation and image obtainment. We thank the Structural Biology Laboratory (SBL) for assistance with cryo-EM grid preparation and screening. Cryo-EM data were collected at the University of Texas Southwestern Medical Center Cryo-Electron Microscopy Facility, which is funded in part by the CPRIT Core Facility Support Award RP170644. We thank SBL and BioHPC for providing computational resources for cryo-EM data processing. This work was funded by the Welch Foundation grant I-1561 (to K.O.), Once Upon a Time… Foundation (to K.O.) and National Institutes of Health grant R01 GM115188 (to K.O.). K.O. is a W. W. Caruth, Jr, Biomedical Scholar with an Earl A. Forsythe Chair in Biomedical Science. D.R.T. is an Effie Marie Scholar. Publisher Copyright: {\textcopyright} 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2020",
month = jun,
day = "1",
doi = "10.1038/s41594-020-0429-1",
language = "English (US)",
volume = "27",
pages = "589--597",
journal = "Nature Structural and Molecular Biology",
issn = "1545-9993",
publisher = "Nature Publishing Group",
number = "6",
}