A Type VI Secretion System Effector Protein, VgrG1, from Aeromonas hydrophila That Induces Host Cell Toxicity by ADP Ribosylation of Actin
10.1128/jb.01260-09
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT We recently delineated the importance of a type VI secretion system (T6SS) gene cluster in the virulence of diarrheal isolate SSU of Aeromonas hydrophila and showed that VasH, a σ 54 activator and T6SS component, was involved in the production of its associated effectors, e.g., hemolysin-coregulated protein. To identify additional T6SS effectors and/or secreted proteins, we subjected culture supernatants from deletion mutants of A. hydrophila , namely, a Δ act mutant (a T2SS-associated cytotoxic enterotoxin-encoding gene) and a Δ act Δ vasH mutant, to 2-dimensional gel electrophoresis and mass spectrometric analysis. Based on these approaches, we identified a member of the VgrG protein family, VgrG1, that contained a vegetative insecticidal protein (VIP-2) domain at its carboxyl-terminal end. Consequently, the vgrG1 gene was cloned in pBI-EGFP and pET-30a vectors to be expressed in HeLa Tet-Off cells and Escherichia coli , respectively. We assessed the ADP-ribosyltransferase (ADPRT) activity of various domains of purified recombinant VgrG1 (rVgrG1) and provided evidence that only the full-length VgrG1, as well as its carboxyl-terminal domain encoding the VIP-2 domain, showed ADPRT activity. Importantly, bacterium-host cell interaction was needed for the T6SS to induce cytotoxicity in eukaryotic cells, and we demonstrated translocation of VgrG1. Furthermore, our data indicated that expression of the genes encoding the full-length VgrG1 and its carboxyl-terminal domain in HeLa Tet-Off cells disrupted the actin cytoskeleton, which was followed by a decrease in cell viability and an increase in apoptosis. Taken together, these findings demonstrated for the first time that VgrG1 of A. hydrophila possessed actin ADPRT activity associated with its VIP-2 domain and that this domain alone was able to induce a rounded phenotype in HeLa Tet-Off cells, followed by apoptosis mediated by caspase 9 activation.

Bibliography

Suarez, G., Sierra, J. C., Erova, T. E., Sha, J., Horneman, A. J., & Chopra, A. K. (2010). A Type VI Secretion System Effector Protein, VgrG1, from Aeromonas hydrophila That Induces Host Cell Toxicity by ADP Ribosylation of Actin. Journal of Bacteriology, 192(1), 155–168.

Authors 6
  1. G. Suarez (first)
  2. J. C. Sierra (additional)
  3. T. E. Erova (additional)
  4. J. Sha (additional)
  5. A. J. Horneman (additional)
  6. A. K. Chopra (additional)
References 53 Referenced 167
  1. Aktories, K., M. Barmann, I. Ohishi, S. Tsuyama, K. H. Jakobs, and E. Habermann. 1986. Botulinum C2 toxin ADP-ribosylates actin. Nature 322 : 390-392. (10.1038/322390a0) / Nature (1986)
  2. Aktories, K., and A. Wegner. 1989. ADP-ribosylation of actin by clostridial toxins. J. Cell Biol. 109 : 1385-1387. (10.1083/jcb.109.4.1385) / J. Cell Biol. (1989)
  3. Aktories, K., U. Weller, and G. S. Chhatwal. 1987. Clostridium botulinum type C produces a novel ADP-ribosyltransferase distinct from botulinum C2 toxin. FEBS Lett. 212 : 109-113. (10.1016/0014-5793(87)81566-1) / FEBS Lett. (1987)
  4. 10.1128/JCM.38.10.3785-3790.2000
  5. 10.1128/MMBR.68.3.373-402.2004
  6. Bingle, L. E., C. M. Bailey, and M. J. Pallen. 2008. Type VI secretion: a beginner's guide. Curr. Opin. Microbiol. 11 : 3-8. (10.1016/j.mib.2008.01.006) / Curr. Opin. Microbiol. (2008)
  7. Cascales, E. 2008. The type VI secretion toolkit. EMBO Rep. 9 : 735-741. (10.1038/embor.2008.131) / EMBO Rep. (2008)
  8. Das, S., and K. Chaudhuri. 2003. Identification of a unique IAHP (IcmF associated homologous proteins) cluster in Vibrio cholerae and other proteobacteria through in silico analysis. In Silico Biol. 3 : 287-300. / In Silico Biol. (2003)
  9. Dudley, E. G., N. R. Thomson, J. Parkhill, N. P. Morin, and J. P. Nataro. 2006. Proteomic and microarray characterization of the AggR regulon identifies a pheU pathogenicity island in enteroaggregative Escherichia coli. Mol. Microbiol. 61 : 1267-1282. (10.1111/j.1365-2958.2006.05281.x) / Mol. Microbiol. (2006)
  10. Edberg, S. C., F. A. Browne, and M. J. Allen. 2007. Issues for microbial regulation: Aeromonas as a model. Crit. Rev. Microbiol. 33 : 89-100. (10.1080/10408410601172180) / Crit. Rev. Microbiol. (2007)
  11. 10.1128/IAI.00487-08
  12. 10.1099/mic.0.2008/016840-0
  13. Frisch, S. M., and H. Francis. 1994. Disruption of epithelial cell-matrix interactions induces apoptosis. J. Cell Biol. 124 : 619-626. (10.1083/jcb.124.4.619) / J. Cell Biol. (1994)
  14. Galindo, C. L., A. A. Fadl, J. Sha, C. Gutierrez, Jr., V. L. Popov, I. Boldogh, B. B. Aggarwal, and A. K. Chopra. 2004. Aeromonas hydrophila cytotoxic enterotoxin activates mitogen-activated protein kinases and induces apoptosis in murine macrophages and human intestinal epithelial cells. J. Biol. Chem. 279 : 37597-37612. (10.1074/jbc.M404641200) / J. Biol. Chem. (2004)
  15. Galindo, C. L., J. Sha, A. A. Fadl, L. Pillai, and A. K. Chopra. 2006. Host immune responses to Aeromonas virulence factors. Curr. Immunol. Rev. 2 : 13-26. (10.2174/157339506775471910) / Curr. Immunol. Rev. (2006)
  16. Gill, D. M., and R. Meren. 1978. ADP-ribosylation of membrane proteins catalyzed by cholera toxin: basis of the activation of adenylate cyclase. Proc. Natl. Acad. Sci. U. S. A. 75 : 3050-3054. (10.1073/pnas.75.7.3050) / Proc. Natl. Acad. Sci. U. S. A. (1978)
  17. Gilmore, A. P., A. D. Metcalfe, L. H. Romer, and C. H. Streuli. 2000. Integrin-mediated survival signals regulate the apoptotic function of Bax through its conformation and subcellular localization. J. Cell Biol. 149 : 431-446. (10.1083/jcb.149.2.431) / J. Cell Biol. (2000)
  18. Han, S., J. A. Craig, C. D. Putnam, N. B. Carozzi, and J. A. Tainer. 1999. Evolution and mechanism from structures of an ADP-ribosylating toxin and NAD complex. Nat. Struct. Biol. 6 : 932-936. (10.1038/13300) / Nat. Struct. Biol. (1999)
  19. 10.1128/IAI.00651-08
  20. Holbourn, K. P., C. C. Shone, and K. R. Acharya. 2006. A family of killer toxins. Exploring the mechanism of ADP-ribosylating toxins. FEBS J. 273 : 4579-4593. (10.1111/j.1742-4658.2006.05442.x) / FEBS J. (2006)
  21. Katada, T., and M. Ui. 1982. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc. Natl. Acad. Sci. U. S. A. 79 : 3129-3133. (10.1073/pnas.79.10.3129) / Proc. Natl. Acad. Sci. U. S. A. (1982)
  22. Koya, R. C., H. Fujita, S. Shimizu, M. Ohtsu, M. Takimoto, Y. Tsujimoto, and N. Kuzumaki. 2000. Gelsolin inhibits apoptosis by blocking mitochondrial membrane potential loss and cytochrome c release. J. Biol. Chem. 275 : 15343-15349. (10.1074/jbc.275.20.15343) / J. Biol. Chem. (2000)
  23. Leiman, P. G., M. Basler, U. A. Ramagopal, J. B. Bonanno, J. M. Sauder, S. Pukatzki, S. K. Burley, S. C. Almo, and J. J. Mekalanos. 2009. Type VI secretion apparatus and phage tail-associated protein complexes share a common evolutionary origin. Proc. Natl. Acad. Sci. U. S. A. 106 : 4154-4159. (10.1073/pnas.0813360106) / Proc. Natl. Acad. Sci. U. S. A. (2009)
  24. Ma, A. T., S. McAuley, S. Pukatzki, and J. J. Mekalanos. 2009. Translocation of a Vibrio cholerae type VI secretion effector requires bacterial endocytosis by host cells. Cell Host Microbe 5 : 234-243. (10.1016/j.chom.2009.02.005) / Cell Host Microbe (2009)
  25. Marchler-Bauer, A., J. B. Anderson, M. K. Derbyshire, C. DeWeese-Scott, N. R. Gonzales, M. Gwadz, L. Hao, S. He, D. I. Hurwitz, J. D. Jackson, Z. Ke, D. Krylov, C. J. Lanczycki, C. A. Liebert, C. Liu, F. Lu, S. Lu, G. H. Marchler, M. Mullokandov, J. S. Song, N. Thanki, R. A. Yamashita, J. J. Yin, D. Zhang, and S. H. Bryant. 2007. CDD: a conserved domain database for interactive domain family analysis. Nucleic Acids Res. 35 : D237-D240. (10.1093/nar/gkl951) / Nucleic Acids Res. (2007)
  26. 10.1128/MCB.21.19.6529-6536.2001
  27. 10.1126/science.1128393
  28. Ohtsu, M., N. Sakai, H. Fujita, M. Kashiwagi, S. Gasa, S. Shimizu, Y. Eguchi, Y. Tsujimoto, Y. Sakiyama, K. Kobayashi, and N. Kuzumaki. 1997. Inhibition of apoptosis by the actin-regulatory protein gelsolin. EMBO J. 16 : 4650-4656. (10.1093/emboj/16.15.4650) / EMBO J. (1997)
  29. Pallen, M. J., R. R. Chaudhuri, and I. R. Henderson. 2003. Genomic analysis of secretion systems. Curr. Opin. Microbiol. 6 : 519-527. (10.1016/j.mib.2003.09.005) / Curr. Opin. Microbiol. (2003)
  30. Pell, L. G., V. Kanelis, L. W. Donaldson, P. L. Howell, and A. R. Davidson. 2009. The phage lambda major tail protein structure reveals a common evolution for long-tailed phages and the type VI bacterial secretion system. Proc. Natl. Acad. Sci. U. S. A. 106 : 4160-4165. (10.1073/pnas.0900044106) / Proc. Natl. Acad. Sci. U. S. A. (2009)
  31. Popoff, M. R., and P. Boquet. 1988. Clostridium spiroforme toxin is a binary toxin which ADP-ribosylates cellular actin. Biochem. Biophys. Res. Commun. 152 : 1361-1368. (10.1016/S0006-291X(88)80435-2) / Biochem. Biophys. Res. Commun. (1988)
  32. 10.1128/iai.56.9.2299-2306.1988
  33. Potvin, E., F. Sanschagrin, and R. C. Levesque. 2008. Sigma factors in Pseudomonas aeruginosa. FEMS Microbiol. Rev. 32 : 38-55. (10.1111/j.1574-6976.2007.00092.x) / FEMS Microbiol. Rev. (2008)
  34. Pukatzki, S., A. T. Ma, A. T. Revel, D. Sturtevant, and J. J. Mekalanos. 2007. Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc. Natl. Acad. Sci. U. S. A. 104 : 15508-15513. (10.1073/pnas.0706532104) / Proc. Natl. Acad. Sci. U. S. A. (2007)
  35. Pukatzki, S., A. T. Ma, D. Sturtevant, B. Krastins, D. Sarracino, W. C. Nelson, J. F. Heidelberg, and J. J. Mekalanos. 2006. Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc. Natl. Acad. Sci. U. S. A. 103 : 1528-1533. (10.1073/pnas.0510322103) / Proc. Natl. Acad. Sci. U. S. A. (2006)
  36. Rao, P. S., Y. Yamada, Y. P. Tan, and K. Y. Leung. 2004. Use of proteomics to identify novel virulence determinants that are required for Edwardsiella tarda pathogenesis. Mol. Microbiol. 53 : 573-586. (10.1111/j.1365-2958.2004.04123.x) / Mol. Microbiol. (2004)
  37. Ribardo, D. A., K. R. Kuhl, I. Boldogh, J. W. Peterson, C. W. Houston, and A. K. Chopra. 2002. Early cell signaling by the cytotoxic enterotoxin of Aeromonas hydrophila in macrophages. Microb. Pathog. 32 : 149-163. (10.1006/mpat.2001.0490) / Microb. Pathog. (2002)
  38. 10.1128/JB.00621-06
  39. 10.1128/IAI.72.10.5983-5992.2004
  40. 10.1128/IAI.70.4.1924-1935.2002
  41. 10.1128/IAI.73.10.6446-6457.2005
  42. Sha, J., S. F. Wang, G. Suarez, J. C. Sierra, A. A. Fadl, T. E. Erova, S. M. Foltz, B. K. Khajanchi, A. Silver, J. Graf, C. H. Schein, and A. K. Chopra. 2007. Further characterization of a type III secretion system (T3SS) and of a new effector protein from a clinical isolate of Aeromonas hydrophila—Part I. Microb. Pathog. 43 : 127-146. (10.1016/j.micpath.2007.05.002) / Microb. Pathog. (2007)
  43. Sierra, J. C., G. Suarez, J. Sha, S. M. Foltz, V. L. Popov, C. L. Galindo, H. R. Garner, and A. K. Chopra. 2007. Biological characterization of a new type III secretion system effector from a clinical isolate of Aeromonas hydrophila—Part II. Microb. Pathog. 43 : 147-160. (10.1016/j.micpath.2007.05.003) / Microb. Pathog. (2007)
  44. Suarez, G., J. C. Sierra, J. Sha, S. Wang, T. E. Erova, A. A. Fadl, S. M. Foltz, A. J. Horneman, and A. K. Chopra. 2008. Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. Microb. Pathog. 44 : 344-361. (10.1016/j.micpath.2007.10.005) / Microb. Pathog. (2008)
  45. Tsuge, H., M. Nagahama, M. Oda, S. Iwamoto, H. Utsunomiya, V. E. Marquez, N. Katunuma, M. Nishizawa, and J. Sakurai. 2008. Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens iota-toxin. Proc. Natl. Acad. Sci. U. S. A. 105 : 7399-7404. (10.1073/pnas.0801215105) / Proc. Natl. Acad. Sci. U. S. A. (2008)
  46. Vandekerckhove, J., B. Schering, M. Barmann, and K. Aktories. 1987. Clostridium perfringens iota toxin ADP-ribosylates skeletal muscle actin in Arg-177. FEBS Lett. 225 : 48-52. (10.1016/0014-5793(87)81129-8) / FEBS Lett. (1987)
  47. Van Ness, B. G., J. B. Howard, and J. W. Bodley. 1980. ADP-ribosylation of elongation factor 2 by diphtheria toxin. Isolation and properties of the novel ribosyl-amino acid and its hydrolysis products. J. Biol. Chem. 255 : 10717-10720. (10.1016/S0021-9258(19)70366-4) / J. Biol. Chem. (1980)
  48. 10.1128/JB.180.16.4102-4110.1998
  49. White, S. R., P. Williams, K. R. Wojcik, S. Sun, P. S. Hiemstra, K. F. Rabe, and D. R. Dorscheid. 2001. Initiation of apoptosis by actin cytoskeletal derangement in human airway epithelial cells. Am. J. Respir. Cell Mol. Biol. 24 : 282-294. (10.1165/ajrcmb.24.3.3995) / Am. J. Respir. Cell Mol. Biol. (2001)
  50. 10.1128/iai.64.1.283-289.1996
  51. Wilson, B. A., and R. J. Collier. 1992. Diphtheria toxin and Pseudomonas aeruginosa exotoxin A: active-site structure and enzymic mechanism. Curr. Top. Microbiol. Immunol. 175 : 27-41. / Curr. Top. Microbiol. Immunol. (1992)
  52. Wolter, K. G., Y. T. Hsu, C. L. Smith, A. Nechushtan, X. G. Xi, and R. J. Youle. 1997. Movement of Bax from the cytosol to mitochondria during apoptosis. J. Cell Biol. 139 : 1281-1292. (10.1083/jcb.139.5.1281) / J. Cell Biol. (1997)
  53. 10.1128/AEM.71.8.4469-4477.2005
Dates
Type When
Created 15 years, 9 months ago (Oct. 30, 2009, 9:38 p.m.)
Deposited 4 years ago (July 29, 2021, 1:13 p.m.)
Indexed 1 month ago (July 26, 2025, 4:50 a.m.)
Issued 15 years, 7 months ago (Jan. 1, 2010)
Published 15 years, 7 months ago (Jan. 1, 2010)
Published Print 15 years, 7 months ago (Jan. 1, 2010)
Funders 0

None

@article{Suarez_2010, title={A Type VI Secretion System Effector Protein, VgrG1, from Aeromonas hydrophila That Induces Host Cell Toxicity by ADP Ribosylation of Actin}, volume={192}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.01260-09}, DOI={10.1128/jb.01260-09}, number={1}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Suarez, G. and Sierra, J. C. and Erova, T. E. and Sha, J. and Horneman, A. J. and Chopra, A. K.}, year={2010}, month=jan, pages={155–168} }