The Helicobacter pylori vacuolating cytotoxin (VacA) is an intracellular, mitochondrial-targeting exotoxin that rapidly causes mitochondrial dysfunction and fragmentation. Although VacA targeting of mitochondria has been reported to alter overall cellular metabolism, there is little known about the consequences of extended exposure to the toxin. Here, we describe studies to address this gap in knowledge, which have revealed that mitochondrial dysfunction and fragmentation are followed by a time-dependent recovery of mitochondrial structure, mitochondrial transmembrane potential, and cellular ATP levels. Cells exposed to VacA also initially demonstrated a reduction in oxidative phosphorylation, as well as increase in compensatory aerobic glycolysis. These metabolic alterations were reversed in cells with limited toxin exposure, congruent with the recovery of mitochondrial transmembrane potential and the absence of cytochrome c release from the mitochondria. Taken together, these results are consistent with a model that mitochondrial structure and function are restored in VacA-intoxicated cells.
Vakifahmetoglu-Norberg, H., Ouchida, A.T. and Norberg, E. (2017) The Role of Mitochondria in Metabolism and Cell Death. Biochemical and Biophysical Research Communications, 482, 426-431. https://doi.org/10.1016/j.bbrc.2016.11.088
Tiku, V., Tan, M.W. and Dikic, I. (2020) Mitochondrial Functions in Infection and Immunity. Trends in Cell Biology, 30, 263-275. https://doi.org/10.1016/j.tcb.2020.01.006
Dodson, M.W. and Guo, M. (2007) Pink1, Parkin, Dj-1 and Mitochondrial Dysfunction in Parkinson’s Disease. Current Opinion in Neurobiology, 17, 331-337. https://doi.org/10.1016/j.conb.2007.04.010
Sharma, L.K., Tiwari, M., Rai, N.K. and Bai, Y. (2019) Mitophagy Activation Repairs Leber’s Hereditary Optic Neuropathy-Associated Mitochondrial Dysfunction and Improves Cell Survival. Human Molecular Genetics, 28, 422-433. https://doi.org/10.1093/hmg/ddy354
Vyas, S., Zaganjor, E. and Haigis, M.C. (2016) Mitochondria and Cancer. Cell, 166, 555-566. https://doi.org/10.1016/j.cell.2016.07.002
Marchi, S., Morroni, G., Pinton, P. and Galluzzi, L. (2022) Control of Host Mitochondria by Bacterial Pathogens. Trends in Microbiology, 30, 452-465. https://doi.org/10.1016/j.tim.2021.09.010
Fielden, L.F., Kang, Y., Newton, H.J. and Stojanovski, D. (2017) Targeting Mitochondria: How Intravacuolar Bacterial Pathogens Manipulate Mitochondria. Cell and Tissue Research, 367, 141-154. https://doi.org/10.1007/s00441-016-2475-x
Howie, H.L., Shiflett, S.L. and So, M. (2008) Extracellular Signal-Regulated Kinase Activation by Neisseria Gonorrhoeae Downregulates Epithelial Cell Proapoptotic Proteins Bad and Bim. Infection and Immunity, 76, 2715-2721. https://doi.org/10.1128/IAI.00153-08
Muller, A., Gunther, D., Brinkmann, V., Hurwitz, R., Meyer, T.F. and Rudel, T. (2000) Targeting of the Pro-Apoptotic Vdac-Like Porin (Porb) of Neisseria Gonorrhoeae to Mitochondria of Infected Cells. The EMBO Journal, 19, 5332-5343. https://doi.org/10.1093/emboj/19.20.5332
Nougayrede, J.P. and Donnenberg, M.S. (2004) Enteropathogenic Escherichia coli Espf Is Targeted to Mitochondria and Is Required to Initiate the Mitochondrial Death Pathway. Cellular Microbiology, 6, 1097-1111. https://doi.org/10.1111/j.1462-5822.2004.00421.x
Yoshizumi, T., Ichinohe, T., Sasaki, O., Otera, H., Kawabata, S.I., Mihara, K. and Koshiba, T. (2014) Influenza a Virus Protein Pb1-F2 Translocates into Mitochondria via Tom40 Channels and Impairs Innate Immunity. Nature Communications, 5, Article No. 4713. https://doi.org/10.1038/ncomms5713
Li, X.D., Sun, L., Seth, R.B., Pineda, G. and Chen, Z.J. (2005) Hepatitis C Virus Protease Ns3/4a Cleaves Mitochondrial Antiviral Signaling Protein off the Mitochondria to Evade Innate Immunity. Proceedings of the National Academy of Sciences of the United States of America, 102, 17717-17722.
Suzuki, M., Danilchanka, O. and Mekalanos, J.J. (2014) Vibrio cholerae T3ss Effector VopE Modulates Mitochondrial Dynamics and Innate Immune Signaling by Targeting Miro Gtpases. Cell Host & Microbe, 16, 581-591. https://doi.org/10.1016/j.chom.2014.09.015
Blanke, S.R. (2005) Micro-Managing the Executioner: Pathogen Targeting of Mitochondria. Trends in Microbiology, 13, 64-71. https://doi.org/10.1016/j.tim.2004.12.007
Hooi, J.K.Y., Lai, W.Y., Ng, W.K., Suen, M.M.Y., Underwood, F.E., Tanyingoh, D., Malfertheiner, P., Graham, D.Y., Wong, V.W.S., Wu, J.C.Y., Chan, F.K.L., Sung, J.J.Y., Kaplan, G.G. and Ng, S.C. (2017) Global Prevalence of Helicobacter pylori Infection: Systematic Review and Meta-Analysis. Gastroenterology, 153, 420-429. https://doi.org/10.1053/j.gastro.2017.04.022
Mahmud, S.A., Qureshi, M.A. and Pellegrino, M.W. (2022) On the Offense and Defense: Mitochondrial Recovery Programs Amidst Targeted Pathogenic Assault. The FEBS Journal, 289, 7014-7037. https://doi.org/10.1111/febs.16126
Correa, P. (1988) A Human Model of Gastric Carcinogenesis. Cancer Research, 48, 3554-3560.
Seeger, A.Y., Ringling, M.D., Zohair, H. and Blanke, S.R. (2020) Risk Factors Associated with Gastric Malignancy during Chronic Helicobacter pylori Infection. Medical Research Archives, 8, 1-20. https://doi.org/10.18103/mra.v8i3.2068
Atherton, J.C., Cao, P., Peek, R.M., Tummuru, M.K.R., Blaser, M.J. and Cover, T.L. (1995) Mosaicism in Vacuolating Cytotoxin Alleles of Helicobacter pylori. Journal of Biological Chemistry, 270, 17771-17777. https://doi.org/10.1074/jbc.270.30.17771
Atherton, J.C., Peek, Jr., Tham, K.T., Cover, T.L. and Blaser, M.J. (1997) Clinical and Pathological Importance of Heterogeneity in Vaca, the Vacuolating Cytotoxin Gene of Helicobacter pylori. Gastroenterology, 112, 92-99. https://doi.org/10.1016/S0016-5085(97)70223-3
Figueiredo, C., Machado, J.C., Pharoah, P., Seruca, R., Sousa, S., Carvalho, R., Capelinha, A.F., Quint, W., Caldas, C., van Doorn, L.J., Carneiro, F. and Sobrinho-Simões, M. (2002) Helicobacter pylori and Interleukin 1 Genotyping: An Opportunity to Identify High-Risk Individuals for Gastric Carcinoma. JNCI: Journal of the National Cancer Institute, 94, 1680-1687. https://doi.org/10.1093/jnci/94.22.1680
Burucoa, C. and Axon, A. (2017) Epidemiology of Helicobacter pylori Infection. Helicobacter, 22, e12403. https://doi.org/10.1111/hel.12403
Salama, N.R., Otto, G., Tompkins, L. and Falkow, S. (2001) Vacuolating Cytotoxin of Helicobacter pylori Plays a Role during Colonization in a Mouse Model of Infection. Infection and Immunity, 69, 730-736. https://doi.org/10.1128/IAI.69.2.730-736.2001
Altobelli, A., Bauer, M., Velez, K., Cover Timothy, L. and Müller, A. (2019) Helicobacter pylori VacA Targets Myeloid Cells in the Gastric Lamina Propria to Promote Peripherally Induced Regulatory T-Cell Differentiation and Persistent Infection. mBio, 10, e00261-19. https://doi.org/10.1128/mBio.00261-19
Tombola, F., Carlesso, C., Szabò, I., de Bernard, M., Reyrat, J.M., Telford, J.L., Rappuoli, R., Montecucco, C., Papini, E. and Zoratti, M. (1999) Helicobacter pylori Vacuolating Toxin Forms Anion-Selective Channels in Planar Lipid Bilayers: Possible Implications for the Mechanism of Cellular Vacuolation. Biophysical Journal, 76, 1401-1409. https://doi.org/10.1016/S0006-3495(99)77301-7
Montecucco, C. and Rappuoli, R. (2001) Living Dangerously: How Helicobacter pylori Survives in the Human Stomach. Nature Reviews Molecular Cell Biology, 2, 457-466. https://doi.org/10.1038/35073084
Willhite, D.C., Cover, T.L. and Blanke, S.R. (2003) Cellular Vacuolation and Mitochondrial Cytochrome c Release Are Independent Outcomes of Helicobacter pylori Vacuolating Cytotoxin Activity That Are Each Dependent on Membrane Channel Formation. Journal of Biological Chemistry, 278, 48204-48209. https://doi.org/10.1074/jbc.M304131200
Willhite, D.C. and Blanke, S.R. (2004) Helicobacter pylori Vacuolating Cytotoxin Enters Cells, Localizes to the Mitochondria, and Induces Mitochondrial Membrane Permeability Changes Correlated to Toxin Channel Activity. Cellular Microbiology, 6, 143-154. https://doi.org/10.1046/j.1462-5822.2003.00347.x
Kimura, M., Goto, S., Wada, A., Yahiro, K., Niidome, T., Hatakeyama, T., Aoyagi, H., Hirayama, T. and Kondo, T. (1999) Vacuolating Cytotoxin Purified from Helicobacter pylori Causes Mitochondrial Damage in Human Gastric Cells. Microbial Pathogenesis, 26, 45-52. https://doi.org/10.1006/mpat.1998.0241
Domańska, G., Motz, C., Meinecke, M., Harsman, A., Papatheodorou, P., Reljic, B., Dian-Lothrop, E.A., Galmiche, A., Kepp, O., Becker, L., Günnewig, K., Wagner, R. and Rassow, J. (2010) Helicobacter pylori VacA Toxin/Subunit P34: Targeting of an Anion Channel to the Inner Mitochondrial Membrane. PLOS Pathogens, 6, e1000878. https://doi.org/10.1371/journal.ppat.1000878
Galmiche, A., Rassow, J., Doye, A., Cagnol, S., Chambard, J.C., Contamin, S., de Thillot, V., Just, I., Ricci, V., Solcia, E., Van Obberghen, E. and Boquet, P. (2000) The N-Terminal 34 kDa Fragment of Helicobacter pylori Vacuolating Cytotoxin Targets Mitochondria and Induces Cytochrome C Release. The EMBO Journal, 19, 6361-6370. https://doi.org/10.1093/emboj/19.23.6361
Terebiznik, M.R., Raju, D., Vázquez, C.L., Torbricki, K., Kulkarni, R., Blanke, S.R., Yoshimori, T., Colombo, M.I. and Jones, N.L. (2009) Effect of Helicobacter pylori’s Vacuolating Cytotoxin on the Autophagy Pathway in Gastric Epithelial Cells. Autophagy, 5, 370-379. https://doi.org/10.4161/auto.5.3.7663
Yahiro, K., Satoh, M., Nakano, M., Hisatsune, J., Isomoto, H., Sap, J., Suzuki, H., Nomura, F., Noda, M., Moss, J. and Hirayama, T. (2012) Low-Density Lipoprotein Receptor-Related Protein-1 (Lrp1) Mediates Autophagy and Apoptosis Caused by Helicobacter pylori Vaca. Journal of Biological Chemistry, 287, 31104-31115. https://doi.org/10.1074/jbc.M112.387498
Raju, D., Hussey, S., Ang, M., Terebiznik, M.R., Sibony, M., Galindo-Mata, E., Gupta, V., Blanke, S.R., Delgado, A., Romero-Gallo, J., Ramjeet, M.S., Mascarenhas, H., Peek, R.M., Correa, P., Streutker, C., Hold, G., Kunstmann, E., Yoshimori, T., Silverberg, M.S., Girardin, S.E., Philpott, D.J., El Omar, E. and Jones, N.L. (2012) Vacuolating Cytotoxin and Variants in Atg16l1 That Disrupt Autophagy Promote Helicobacter pylori Infection in Humans. Gastroenterology, 142, 1160-1171. https://doi.org/10.1053/j.gastro.2012.01.043
Kuck, D., Kolmerer, B., Iking-Konert, C., Krammer, P.H., Stremmel, W. and Rudi, J. (2001) Vacuolating Cytotoxin of Helicobacter pylori Induces Apoptosis in the Human Gastric Epithelial Cell Line AGS. Infection and Immunity, 69, 5080-5087. https://doi.org/10.1128/IAI.69.8.5080-5087.2001
Cover, T.L., Krishna, U.S., Israel, D.A. and Peek, R.M. (2003) Induction of Gastric Epithelial Cell Apoptosis by Helicobacter pylori Vacuolating Cytotoxin. Cancer Research, 63, 951-957.
Radin, J.N., González-Rivera, C., Ivie, S.E., McClain, M.S. and Cover, T.L. (2011) Helicobacter pylori VacA Induces Programmed Necrosis in Gastric Epithelial Cells. Infection and Immunity, 79, 2535-2543. https://doi.org/10.1128/IAI.01370-10
Garner, J.A. and Cover, T.L. (1996) Binding and Internalization of the Helicobacter pylori Vacuolating Cytotoxin by Epithelial Cells. Infection and Immunity, 64, 4197-4203. https://doi.org/10.1128/iai.64.10.4197-4203.1996
Gauthier, N.C., Monzo, P., Gonzalez, T., Doye, A., Oldani, A., Gounon, P., Ricci, V., Cormont, M. and Boquet, P. (2007) Early Endosomes Associated with Dynamic F-Actin Structures Are Required for Late Trafficking of H. pylori VacA Toxin. Journal of Cell Biology, 177, 343-354. https://doi.org/10.1083/jcb.200609061
Gauthier, N.C., Monzo, P., Kaddai, V., Doye, A., Ricci, V. and Boquet, P. (2005) Helicobacter pylori VacA Cytotoxin: A Probe for a Clathrin-Independent and Cdc42-Dependent Pinocytic Pathway Routed to Late Endosomes. Molecular Biology of the Cell, 16, 4852-4866. https://doi.org/10.1091/mbc.e05-05-0398
Gupta, V.R., Patel, H.K., Kostolansky, S.S., Ballivian, R.A., Eichberg, J. and Blanke, S.R. (2008) Sphingomyelin Functions as a Novel Receptor for Helicobacter pylori VacA. PLOS Pathogens, 4, e1000073. https://doi.org/10.1371/journal.ppat.1000073
Gupta, V.R., Wilson, B.A. and Blanke, S.R. (2010) Sphingomyelin Is Important for the Cellular Entry and Intracellular Localization of Helicobacter pylori VacA. Cellular Microbiology, 12, 1517-1533. https://doi.org/10.1111/j.1462-5822.2010.01487.x
Patel, H.K., Willhite, D.C., Patel, R.M., Ye, D., Williams, C.L., Torres, E.M., Marty, K.B., MacDonald, R.A. and Blanke, S.R. (2002) Plasma Membrane Cholesterol Modulates Cellular Vacuolation Induced by the Helicobacter pylori Vacuolating Cytotoxin. Infection and Immunity, 70, 4112-4123. https://doi.org/10.1128/IAI.70.8.4112-4123.2002
Calore, F., Genisset, C., Casellato, A., Rossato, M., Codolo, G., Esposti, M.D., Scorrano, L. and de Bernard, M. (2010) Endosome-Mitochondria Juxtaposition during Apoptosis Induced by H. Pylori VacA. Cell Death and Differentiation, 17, 1707-1716. https://doi.org/10.1038/cdd.2010.42
Foo, J.H., Culvenor, J.G., Ferrero, R.L., Kwok, T., Lithgow, T. and Gabriel, K. (2010) Both the P33 and P55 Subunits of the Helicobacter pylori VacA Toxin Are Targeted to Mammalian Mitochondria. Journal of Molecular Biology, 401, 792-798. https://doi.org/10.1016/j.jmb.2010.06.065
Jain, P., Luo, Z.Q. and Blanke, S.R. (2011) Helicobacter pylori Vacuolating Cytotoxin a (VacA) Engages the Mitochondrial Fission Machinery to Induce Host Cell Death. Proceedings of the National Academy of Sciences of the United States of America, 108, 16032-16037.
Kim, I.J., Lee, J., Oh, S.J., Yoon, M.S., Jang, S.S., Holland, R.L., Reno, M.L., Hamad, M.N., Maeda, T., Chung, H.J., Chen, J. and Blanke, S.R. (2018) Helicobacter pylori Infection Modulates Host Cell Metabolism through VacA-Dependent Inhibition of Mtorc1. Cell Host & Microbe, 23, 583-593.E8. https://doi.org/10.1016/j.chom.2018.04.006
Wu, S., Zhou, F., Zhang, Z. and Xing, D. (2011) Mitochondrial Oxidative Stress Causes Mitochondrial Fragmentation Via Differential Modulation of Mitochondrial Fission-Fusion Proteins. FEBS Journal, 278, 941-954. https://doi.org/10.1111/j.1742-4658.2011.08010.x
Youle, R.J. and van der Bliek, A.M. (2012) Mitochondrial Fission, Fusion, and Stress. Science, 337, 1062-1065. https://doi.org/10.1126/science.1219855
Toyama, E.Q., Herzig, S., Courchet, J., Lewis, T.L., Losón, O.C., Hellberg, K., Young, N.P., Chen, H., Polleux, F., Chan, D.C. and Shaw, R.J. (2016) Amp-Activated Protein Kinase Mediates Mitochondrial Fission in Response to Energy Stress. Science, 351, 275-281. https://doi.org/10.1126/science.aab4138
Perdiz, D., Oziol, L. and Poüs, C. (2019) Early Mitochondrial Fragmentation Is a Potential in vitro Biomarker of Environmental Stress. Chemosphere, 223, 577-587. https://doi.org/10.1016/j.chemosphere.2019.02.044
Knott, A.B., Perkins, G., Schwarzenbacher, R. and Bossy-Wetzel, E. (2008) Mitochondrial Fragmentation in Neurodegeneration. Nature Reviews Neuroscience, 9, 505-518. https://doi.org/10.1038/nrn2417
Chang, X., Li, Y., Cai, C., Wu, F., He, J., Zhang, Y., Zhong, J., Tan, Y., Liu, R., Zhu, H. and Zhou, H. (2022) Mitochondrial Quality Control Mechanisms as Molecular Targets in Diabetic Heart. Metabolism, 137, Article ID: 155313. https://doi.org/10.1016/j.metabol.2022.155313
Nakayama, M., Hisatsune, J., Yamasaki, E., Nishi, Y., Wada, A., Kurazono, H., Sap, J., Yahiro, K., Moss, J. and Hirayama, T. (2006) Clustering of Helicobacter pylori VacA in Lipid Rafts, Mediated by Its Receptor, Receptor-Like Protein Tyrosine Phosphatase β, Is Required for Intoxication in Az-521 Cells. Infection and Immunity, 74, 6571-6580. https://doi.org/10.1128/IAI.00356-06
Menaker Rena, J., Ceponis Peter, J.M. and Jones Nicola, L. (2004) Helicobacter pylori Induces Apoptosis of Macrophages in Association with Alterations in the Mitochondrial Pathway. Infection and Immunity, 72, 2889-2898. https://doi.org/10.1128/IAI.72.5.2889-2898.2004
McClain, M.S., Cao, P., Iwamoto, H., Vinion-Dubiel, A.D., Szabo, G., Shao, Z. and Cover, T.L. (2001) A 12-Amino-Acid Segment, Present in Type S2 But Not Type S1 Helicobacter pylori VacA Proteins, Abolishes Cytotoxin Activity and Alters Membrane Channel Formation. Journal of Bacteriology, 183, 6499-6508. https://doi.org/10.1128/JB.183.22.6499-6508.2001
Gauthier, N.C., Ricci, V., Gounon, P., Doye, A., Tauc, M., Poujeol, P. and Boquet, P. (2004) Glycosylphosphatidylinositol-Anchored Proteins and Actin Cytoskeleton Modulate Chloride Transport by Channels Formed by the Helicobacter pylori Vacuolating Cytotoxin VacA in HeLa Cells. Journal of Biological Chemistry, 279, 9481-9489. https://doi.org/10.1074/jbc.M312040200
de Bernard, M., Burroni, D., Papini, E., Rappuoli, R., Telford, J. and Montecucco, C. (1998) Identification of the Helicobacter pylori VacA Toxin Domain Active in the Cell Cytosol. Infection and Immunity, 66, 6014-6016. https://doi.org/10.1128/IAI.66.12.6014-6016.1998
Seto, K., Hayashi-Kuwabara, Y., Yoneta, T., Suda, H. and Tamaki, H. (1998) Vacuolation Induced by Cytotoxin from Helicobacter pylori Is Mediated by the EGF Receptor in Hela Cells. FEBS Letters, 431, 347-350. https://doi.org/10.1016/S0014-5793(98)00788-1
Yahiro, K., Niidome, T., Hatakeyama, T., Aoyagi, H., Kurazono, H., Padilla, P.I., Wada, A. and Hirayama, T. (1997) Helicobacter pylori Vacuolating Cytotoxin Binds to the 140-kDa Protein in Human Gastric Cancer Cell Lines, Az-521 and AGS. Biochemical and Biophysical Research Communications, 238, 629-632. https://doi.org/10.1006/bbrc.1997.7345
Hatefi, Y. (1985) The Mitochondrial Electron Transport and Oxidative Phosphorylation System. Annual Review of Biochemistry, 54, 1015-1069. https://doi.org/10.1146/annurev.bi.54.070185.005055
Lunt, S.Y. and Vander Heiden, M.G. (2011) Aerobic Glycolysis: Meeting the Metabolic Requirements of Cell Proliferation. Annual Review of Cell and Developmental Biology, 27, 441-464. https://doi.org/10.1146/annurev-cellbio-092910-154237
Rabinowitz, J.D. and Enerbäck, S. (2020) Lactate: The Ugly Duckling of Energy Metabolism. Nature Metabolism, 2, 566-571. https://doi.org/10.1038/s42255-020-0243-4
Vaupel, P., Schmidberger, H. and Mayer, A. (2019) The Warburg Effect: Essential Part of Metabolic Reprogramming and Central Contributor to Cancer Progression. International Journal of Radiation Biology, 95, 912-919. https://doi.org/10.1080/09553002.2019.1589653
Foegeding, N.J., Raghunathan, K., Campbell, A.M., Kim, S.W., Lau, K.S., Kenworthy, A.K., Cover, T.L. and Ohi, M.D. (2019) Intracellular Degradation of Helicobacter pylori VacA Toxin as a Determinant of Gastric Epithelial Cell Viability. Infection and Immunity, 87, e00783-18. https://doi.org/10.1128/IAI.00783-18
Lee, H. and Yoon, Y. (2016) Mitochondrial Fission and Fusion. Biochemical Society Transactions, 44, 1725-1735. https://doi.org/10.1042/BST20160129
Marchetti, M., Aricò, B., Burroni, D., Figura, N., Rappuoli, R. and Ghiara, P. (1995) Development of a Mouse Model of Helicobacter pylori Infection That Mimics Human Disease. Science, 267, 1655-1658. https://doi.org/10.1126/science.7886456
Ghiara, P., Marchetti, M., Blaser, M.J., Tummuru, M.K.R., Cover, T.L., Segal, E.D., Tompkins, L.S. and Rappuoli, R. (1995) Role of the Helicobacter pylori Virulence Factors Vacuolating Cytotoxin, CagA, and Urease in a Mouse Model of Disease. Infection and Immunity, 63, 4154-4160. https://doi.org/10.1128/iai.63.10.4154-4160.1995
Fujikawa, A., Shirasaka, D., Yamamoto, S., Ota, H., Yahiro, K., Fukada, M., Shintani, T., Wada, A., Aoyama, N., Hirayama, T., Fukamachi, H. and Noda, M. (2003) Mice Deficient in Protein Tyrosine Phosphatase Receptor Type Z Are Resistant to Gastric Ulcer Induction by VacA of Helicobacter pylori. Nature Genetics, 33, 375-381. https://doi.org/10.1038/ng1112
Xiang, Z., Censini, S., Bayeli, P.F., Telford, J.L., Figura, N., Rappuoli, R. and Covacci, A. (1995) Analysis of Expression of CagA and VacA Virulence Factors in 43 Strains of Helicobacter pylori Reveals That Clinical Isolates Can Be Divided into Two Major Types and That CagA Is Not Necessary for Expression of the Vacuolating Cytotoxin. Infection and Immunity, 63, 94-98. https://doi.org/10.1128/iai.63.1.94-98.1995
Telford, J.L., Ghiara, P., Dell’Orco, M., Comanducci, M., Burroni, D., Bugnoli, M., Tecce, M.F., Censini, S., Covacci, A. and Xiang, Z. (1994) Gene Structure of the Helicobacter pylori Cytotoxin and Evidence of Its Key Role in Gastric Disease. Journal of Experimental Medicine, 179, 1653-1658. https://doi.org/10.1084/jem.179.5.1653
Yamasaki, E., Wada, A., Kumatori, A., Nakagawa, I., Funao, J., Nakayama, M., Hisatsune, J., Kimura, M., Moss, J. and Hirayama, T. (2006) Helicobacter pylori Vacuolating Cytotoxin Induces Activation of the Proapoptotic Proteins Bax and Bak, Leading to Cytochrome C Release and Cell Death, Independent of Vacuolation. Journal of Biological Chemistry, 281, 11250-11259. https://doi.org/10.1074/jbc.M509404200
Akazawa, Y., Isomoto, H., Matsushima, K., Kanda, T., Minami, H., Yamaghchi, N., Taura, N., Shiozawa, K., Ohnita, K., Takeshima, F., Nakano, M., Moss, J., Hirayama, T. and Nakao, K. (2013) Endoplasmic Reticulum Stress Contributes to Helicobacter pylori VacA-Induced Apoptosis. PLOS ONE, 8, e82322. https://doi.org/10.1371/journal.pone.0082322
Yahiro, K., Akazawa, Y., Nakano, M., Suzuki, H., Hisatune, J., Isomoto, H., Sap, J., Noda, M., Moss, J. and Hirayama, T. (2015) Helicobacter pylori VacA Induces Apoptosis by Accumulation of Connexin 43 in Autophagic Vesicles via a Rac1/Erk-Dependent Pathway. Cell Death Discovery, 1, Article No. 15035. https://doi.org/10.1038/cddiscovery.2015.35
Zhu, P., Xue, J., Zhang, Z.J., Jia, Y.P., Tong, Y.N., Han, D., Li, Q., Xiang, Y., Mao, X.H. and Tang, B. (2017) Helicobacter pylori VacA Induces Autophagic Cell Death in Gastric Epithelial Cells via the Endoplasmic Reticulum Stress Pathway. Cell Death & Disease, 8, Article No. 3207. https://doi.org/10.1038/s41419-017-0011-x
Bock, F.J. and Tait, S.W.G. (2020) Mitochondria as Multifaceted Regulators of Cell Death. Nature Reviews Molecular Cell Biology, 21, 85-100. https://doi.org/10.1038/s41580-019-0173-8
Holland, R.L., Bosi, K.D., Harpring, G.H., Luo, J., Wallig, M., Phillips, H. and Blanke, S.R. (2020) Chronic in Vivo Exposure to Helicobacter pylori VacA: Assessing the Efficacy of Automated and Long-Term Intragastric Toxin Infusion. Scientific Reports, 10, Article No. 9307. https://doi.org/10.1038/s41598-020-65787-3