Home LiteratureArticle Details
PMID: 21722286 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Shiga toxins induce autophagy leading to differential signalling pathways in toxin-sensitive and toxin-resistant human cells.

Cellular microbiology ·Vol. 13 ·No. 10 ·2011-10-00 ·Pages 1479-96

Lee MS, Cherla RP, Jenson MH, Leyva-Illades D, Martinez-Moczygemba M, Tesh VL

Abstract

The bacterial virulence factors Shiga toxins (Stxs) are expressed by Shigella dysenteriae serotype 1 and certain Escherichia coli strains. Stxs are protein synthesis inhibitors and induce apoptosis in many cell types. Stxs induce apoptosis via prolonged endoplasmic reticulum stress signalling to activate both extrinsic and intrinsic pathways in human myeloid cells. Studies have shown that autophagy, a lysosome-dependent catabolic process, may be associated with activation of pro-survival or death processes. It is currently unknown if autophagy contributes to apoptosis or protects cells from Stxs. To study cellular responses to Stxs, we intoxicated toxin-sensitive cells (THP-1 and HK-2 cells), and toxin-resistant cells (primary human monocyte-derived macrophages) and examined toxin intracellular trafficking and autophagosome formation. Stxs translocated to different cell compartments in toxin-resistant versus toxin-sensitive cells. Confocal microscopy revealed autophagosome formation in both toxin-resistant and toxin-sensitive cells. Proteolytic cleavage of Atg5 and Beclin-1 plays pivotal roles in switching non-cytotoxic autophagy to cell death signalling. We detected cleaved forms of Atg5 and Beclin-1 in Stx-treated toxin-sensitive cells, while cleaved caspases, calpains, Atg5 and Beclin-1 were not detected in toxin-resistant primary human monocytes and macrophages. These findings suggest that toxin sensitivity correlates with caspase and calpain activation, leading to Atg5 and Beclin-1 cleavage.

MeSH Terms
Apoptosis Regulatory Proteins/metabolism Autophagy Autophagy-Related Protein 5 Beclin-1 Calpain/metabolism Caspases/metabolism Cells, Cultured Escherichia coli/pathogenicity Host-Pathogen Interactions Humans Membrane Proteins/metabolism Microtubule-Associated Proteins/metabolism Shiga Toxin Shiga Toxins/toxicity Shigella dysenteriae/pathogenicity Signal Transduction
Chemicals
ATG5 protein, human Apoptosis Regulatory Proteins Autophagy-Related Protein 5 BECN1 protein, human Beclin-1 Membrane Proteins Microtubule-Associated Proteins Shiga Toxins Shiga Toxin Calpain Caspases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lee Moo-Seung
Department of Microbial and Molecular Pathogenesis, Texas A&M Health Science Center, Bryan, TX 77807, USA.
Cherla Rama P
Jenson Matthew H
Leyva-Illades Dinorah
Martinez-Moczygemba Margarita
Tesh Vernon L
References (66)
66 references, click to expand
  1. Global burden of Shigella infections: implications for vaccine development and implementation of control strategies.
    Bull World Health Organ. 1999;77(8):651-66 PMID: 10516787
  2. Simultaneous induction of apoptotic and survival signaling pathways in macrophage-like THP-1 cells by Shiga toxin 1.
    Infect Immun. 2007 Mar;75(3):1291-302 PMID: 17194804
  3. Comparison of the relative toxicities of Shiga-like toxins type I and type II for mice.
    Infect Immun. 1993 Aug;61(8):3392-402 PMID: 8335369
  4. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to Shiga toxin.
    J Immunol. 2008 Jul 15;181(2):1460-9 PMID: 18606701
  5. Interactions between Shiga toxins and human polymorphonuclear leukocytes.
    J Leukoc Biol. 2008 Oct;84(4):1019-27 PMID: 18625912
  6. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes.
    Proc Natl Acad Sci U S A. 1982 Mar;79(6):1889-92 PMID: 6952238
  7. Double-stranded RNA-activated protein kinase mediates induction of interleukin-8 expression by deoxynivalenol, Shiga toxin 1, and ricin in monocytes.
    Toxicol Sci. 2008 Oct;105(2):322-30 PMID: 18599499
  8. The MAP kinase-activated protein kinase 2 (MK2) contributes to the Shiga toxin-induced inflammatory response.
    Cell Microbiol. 2010 Apr 1;12(4):516-29 PMID: 19951368
  9. Shiga toxin, Shiga-like toxin II variant, and ricin are all single-site RNA N-glycosidases of 28 S RNA when microinjected into Xenopus oocytes.
    J Biol Chem. 1989 Jan 5;264(1):596-601 PMID: 2642481
  10. Shiga toxin B-subunit binds to the chaperone BiP and the nucleolar protein B23.
    Biol Cell. 2006 Feb;98(2):125-34 PMID: 15853775
  11. Autophagy and metabolism.
    Science. 2010 Dec 3;330(6009):1344-8 PMID: 21127245
  12. Switch between apoptosis and autophagy: radiation-induced endoplasmic reticulum stress?
    Cell Cycle. 2007 Apr 1;6(7):793-8 PMID: 17377498
  13. Macropinocytosis in Shiga toxin 1 uptake by human intestinal epithelial cells and transcellular transcytosis.
    Am J Physiol Gastrointest Liver Physiol. 2009 Jan;296(1):G78-92 PMID: 18974311
  14. Shiga toxin 1-induced proinflammatory cytokine production is regulated by the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin signaling pathway.
    Infect Immun. 2009 Sep;77(9):3919-31 PMID: 19596774
  15. The B subunit of Shiga toxin coupled to full-size antigenic protein elicits humoral and cell-mediated immune responses associated with a Th1-dominant polarization.
    Int Immunol. 2003 Oct;15(10):1161-71 PMID: 13679386
  16. Shiga toxin induces tubular membrane invaginations for its uptake into cells.
    Nature. 2007 Nov 29;450(7170):670-5 PMID: 18046403
  17. Site of action of a Vero toxin (VT2) from Escherichia coli O157:H7 and of Shiga toxin on eukaryotic ribosomes. RNA N-glycosidase activity of the toxins.
    Eur J Biochem. 1988 Jan 15;171(1-2):45-50 PMID: 3276522
  18. Shiga toxin receptor glycolipid binding. Pathology and utility.
    Methods Mol Med. 2003;73:165-86 PMID: 12375428
  19. Shiga toxins 1 and 2 translocate differently across polarized intestinal epithelial cells.
    Infect Immun. 1999 Dec;67(12):6670-7 PMID: 10569789
  20. Characterization of non-toxic mutant toxins of Vero toxin 1 that were constructed by replacing amino acids in the A subunit.
    Microb Pathog. 1993 Sep;15(3):169-76 PMID: 8271917
  21. Shiga toxin 1 triggers a ribotoxic stress response leading to p38 and JNK activation and induction of apoptosis in intestinal epithelial cells.
    Infect Immun. 2003 Mar;71(3):1497-504 PMID: 12595468
  22. Retrograde transport of endocytosed Shiga toxin to the endoplasmic reticulum.
    Nature. 1992 Aug 6;358(6386):510-2 PMID: 1641040
  23. Endocytosis and retrograde transport of Shiga toxin.
    Toxicon. 2010 Dec 15;56(7):1181-5 PMID: 19951719
  24. Targeting of Shiga toxin B-subunit to retrograde transport route in association with detergent-resistant membranes.
    Mol Biol Cell. 2001 Aug;12(8):2453-68 PMID: 11514628
  25. Development by self-digestion: molecular mechanisms and biological functions of autophagy.
    Dev Cell. 2004 Apr;6(4):463-77 PMID: 15068787
  26. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture.
    Cancer Commun. 1991 Jul;3(7):207-12 PMID: 1867954
  27. Shiga toxin type 1 activates tumor necrosis factor-alpha gene transcription and nuclear translocation of the transcriptional activators nuclear factor-kappaB and activator protein-1.
    Blood. 1998 Jul 15;92(2):558-66 PMID: 9657756
  28. Functional and physical interaction between Bcl-X(L) and a BH3-like domain in Beclin-1.
    EMBO J. 2007 May 16;26(10):2527-39 PMID: 17446862
  29. Silencing of Bak ameliorates apoptosis of human proximal tubular epithelial cells by Escherichia coli-derived Shiga toxin 2.
    Infection. 2005 Oct;33(5-6):362-7 PMID: 16258868
  30. Autophagy: molecular machinery, regulation, and implications for renal pathophysiology.
    Am J Physiol Renal Physiol. 2009 Aug;297(2):F244-56 PMID: 19279132
  31. Effects of verocytotoxin-1 on nonadherent human monocytes: binding characteristics, protein synthesis, and induction of cytokine release.
    Blood. 1996 Jul 1;88(1):174-83 PMID: 8704172
  32. Toxin-induced cell lysis: protection by 3-methyladenine and cycloheximide.
    Exp Cell Res. 1992 Jun;200(2):253-62 PMID: 1572394
  33. A protein conjugation system essential for autophagy.
    Nature. 1998 Sep 24;395(6700):395-8 PMID: 9759731
  34. Shiga toxin 1 induces apoptosis through the endoplasmic reticulum stress response in human monocytic cells.
    Cell Microbiol. 2008 Mar;10(3):770-80 PMID: 18005243
  35. Impairment by verotoxin of tubular function contributes to the renal damage seen in haemolytic uraemic syndrome.
    J Infect. 1993 Nov;27(3):339-41 PMID: 8308331
  36. Shiga toxins--from cell biology to biomedical applications.
    Nat Rev Microbiol. 2010 Feb;8(2):105-16 PMID: 20023663
  37. Binding and transfer of verocytotoxin by polymorphonuclear leukocytes in hemolytic uremic syndrome.
    Blood. 2000 Jun 1;95(11):3396-402 PMID: 10828021
  38. Activation of Src family kinase yes induced by Shiga toxin binding to globotriaosyl ceramide (Gb3/CD77) in low density, detergent-insoluble microdomains.
    J Biol Chem. 1999 Dec 3;274(49):35278-82 PMID: 10575015
  39. Differentiation-associated toxin receptor modulation, cytokine production, and sensitivity to Shiga-like toxins in human monocytes and monocytic cell lines.
    Infect Immun. 1996 Apr;64(4):1173-80 PMID: 8606075
  40. Self-eating and self-killing: crosstalk between autophagy and apoptosis.
    Nat Rev Mol Cell Biol. 2007 Sep;8(9):741-52 PMID: 17717517
  41. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy.
    Cell. 2005 Sep 23;122(6):927-39 PMID: 16179260
  42. Does autophagy contribute to cell death?
    Autophagy. 2005 Jul;1(2):66-74 PMID: 16874022
  43. Overview of macroautophagy regulation in mammalian cells.
    Cell Res. 2010 Jul;20(7):748-62 PMID: 20548331
  44. Calpain-mediated cleavage of Atg5 switches autophagy to apoptosis.
    Nat Cell Biol. 2006 Oct;8(10):1124-32 PMID: 16998475
  45. Shiga toxin receptor Gb3Cer/CD77: tumor-association and promising therapeutic target in pancreas and colon cancer.
    PLoS One. 2009 Aug 28;4(8):e6813 PMID: 19714252
  46. ZAK: a MAP3Kinase that transduces Shiga toxin- and ricin-induced proinflammatory cytokine expression.
    Cell Microbiol. 2008 Jul;10(7):1468-77 PMID: 18331592
  47. Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network.
    EMBO Rep. 2001 Apr;2(4):330-5 PMID: 11306555
  48. Upregulation of Beclin-1 expression and phosphorylation of Bcl-2 and p53 are involved in the JNK-mediated autophagic cell death.
    Biochem Biophys Res Commun. 2009 May 15;382(4):726-9 PMID: 19318089
  49. Cytoprotective effect of curcumin in human proximal tubule epithelial cells exposed to shiga toxin.
    Biochem Biophys Res Commun. 2001 Apr 27;283(1):36-41 PMID: 11322764
  50. Connecting endoplasmic reticulum stress to autophagy by unfolded protein response and calcium.
    Cell Death Differ. 2007 Sep;14(9):1576-82 PMID: 17612585
  51. Food-related illness and death in the United States.
    Emerg Infect Dis. 1999 Sep-Oct;5(5):607-25 PMID: 10511517
  52. Autophagy and cell death.
    Curr Top Dev Biol. 2007;78:217-45 PMID: 17338918
  53. CNS manifestations of the hemolytic-uremic syndrome. Relationship to metabolic alterations and prognosis.
    Am J Dis Child. 1980 Sep;134(9):869-72 PMID: 7416114
  54. The in vitro cleavage of the hAtg proteins by cell death proteases.
    Autophagy. 2010 Nov;6(8):1042-56 PMID: 21121091
  55. Bcl-2 regulates the onset of shiga toxin 1-induced apoptosis in THP-1 cells.
    Infect Immun. 2009 Dec;77(12):5233-44 PMID: 19752028
  56. Caspase-mediated cleavage of Beclin-1 inactivates Beclin-1-induced autophagy and enhances apoptosis by promoting the release of proapoptotic factors from mitochondria.
    Cell Death Dis. 2010;1:e18 PMID: 21364619
  57. ER stress (PERK/eIF2alpha phosphorylation) mediates the polyglutamine-induced LC3 conversion, an essential step for autophagy formation.
    Cell Death Differ. 2007 Feb;14(2):230-9 PMID: 16794605
  58. Signaling through C/EBP homologous protein and death receptor 5 and calpain activation differentially regulate THP-1 cell maturation-dependent apoptosis induced by Shiga toxin type 1.
    Infect Immun. 2010 Aug;78(8):3378-91 PMID: 20515924
  59. Shiga toxin 1-induced activation of c-Jun NH(2)-terminal kinase and p38 in the human monocytic cell line THP-1: possible involvement in the production of TNF-alpha.
    J Leukoc Biol. 2002 Jan;71(1):107-14 PMID: 11781386
  60. Comparative evaluation of apoptosis induced by Shiga toxin 1 and/or lipopolysaccharides in human monocytic and macrophage-like cells.
    Microb Pathog. 2005 Feb-Mar;38(2-3):63-76 PMID: 15748808
  61. Structure of shiga toxin type 2 (Stx2) from Escherichia coli O157:H7.
    J Biol Chem. 2004 Jun 25;279(26):27511-7 PMID: 15075327
  62. Autophagy is activated for cell survival after endoplasmic reticulum stress.
    Mol Cell Biol. 2006 Dec;26(24):9220-31 PMID: 17030611
  63. Up-regulation of tissue factor activity on human proximal tubular epithelial cells in response to Shiga toxin.
    Kidney Int. 2005 Jun;67(6):2254-66 PMID: 15882267
  64. Shiga toxin-2 triggers endothelial leukocyte adhesion and transmigration via NF-kappaB dependent up-regulation of IL-8 and MCP-1.
    Kidney Int. 2002 Sep;62(3):846-56 PMID: 12164866
  65. Induction of apoptosis by Shiga toxins.
    Future Microbiol. 2010 Mar;5(3):431-53 PMID: 20210553
  66. LC3 conjugation system in mammalian autophagy.
    Int J Biochem Cell Biol. 2004 Dec;36(12):2503-18 PMID: 15325588
Article Info
Journal
Cellular microbiology
Abbr.
Cell Microbiol
ISSN
1462-5822
DOI
10.1111/j.1462-5822.2011.01634.x
Published
2011-10-00
Epub
2011-00-04
Pages
1479-96
Language
English
Region
England
NLM ID
100883691
PMCID
PMC3173519
Subset
IM
Grants
NIAID NIH HHS · R01 AI034530 · United States
NIAID NIH HHS · R01 AI034530-14 · United States
NIAID NIH HHS · R01 AI34530 · United States
External Links
PubMed source DOI: 10.1111/j.1462-5822.2011.01634.x PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2