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Novel nuclear targeting coiled-coil protein of Helicobacter pylori showing Ca2+-independent, Mg2+-dependent DNase I activity

  • Microbial Physiology and Biochemistry
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Abstract

HP0059, an uncharacterized gene of Helicobacter pylori, encodes a 284-aa-long protein containing a nuclear localization sequence (NLS) and multiple leucine-rich heptad repeats. Effects of HP0059 proteins in human stomach cells were assessed by incubation of recombinant HP0059 proteins with the AGS human gastric carcinoma cell line. Wild-type HP0059 proteins showed cytotoxicity in AGS cells in a concentration-dependent manner, whereas NLS mutant protein showed no effect, suggesting that the cytotoxicity is attributed to host nuclear localization. AGS cells transfected with pEGFP-HP0059 plasmid showed strong GFP signal merged to the chromosomal DNA region. The chromosome was fragmented into multiple distinct dots merged with the GFP signal after 12 h of incubation. The chromosome fragmentation was further explored by incubation of AGS chromosomal DNA with recombinant HP0059 proteins, which leaded to complete degradation of the chromosomal DNA. HP0059 protein also degraded circular plasmid DNA without consensus, being an indication of DNase I activity. The DNase was activated by MgCl2, but not by CaCl2. The activity was completely blocked by EDTA. The optimal pH and temperature for DNase activity were 7.0–8.0 and 55°C, respectively. These results indicate that HP0059 possesses a novel DNase I activity along with a role in the genomic instability of human gastric cells, which may result in the transformation of gastric cells.

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References

  • Apostolovic, B., Deacon, S.P., Duncan, R., and Klok, H.A. 2011. Cell uptake and trafficking behavior of non-covalent, coiled-coil based polymer–drug conjugates. Macromol. Rap. Commun. 32, 11–18.

    Article  CAS  Google Scholar 

  • Bae, M., Lim, J.W., and Kim, H. 2013. Oxidative DNA damage response in Helicobacter pylori-infected Mongolian Gerbils. J. Cancer Prev. 18, 271–275.

    Article  PubMed  PubMed Central  Google Scholar 

  • Bitter, W., Houben, E.N., Bottai, D., Brodin, P., Brown, E.J., Cox, J.S., Derbyshire, K., Fortune, S.M., Gao, L.Y., Liu, J., et al. 2009. Systematic genetic nomenclature for type VII secretion systems. PLoS Pathog. 5, e1000507.

    Article  Google Scholar 

  • Brodin, P., Rosenkrands, I., Andersen, P., Cole, S.T., and Brosch, R. 2004. ESAT-6 proteins: protective antigens and virulence factors? Trend. Microbiol. 12, 500–508.

    Article  CAS  Google Scholar 

  • Brown, J.H., Kim, K.H., Jun, G., Greenfield, N.J., Dominguez, R., Volkmann, N., Hitchcock-DeGregori, S.E., and Cohen, C. 2001. Deciphering the design of the tropomyosin molecule. Proc. Natl. Acad. Sci. USA 98, 8496–8501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Censini, S., Lange, C., Xiang, Z., Crabtree, J.E., Ghiara, P., Borodovsky, M., Rappuoli, R., and Covacci, A. 1996. cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and diseaseassociated virulence factors. Proc. Natl. Acad. Sci. USA 93, 14648–14653.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cole, C., Barber, J.D., and Barton, G.J. 2008. The Jpred 3 secondary structure prediction server. Nucleic Acids Res. 36, W197–W201.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cornelis, G.R. and Wolf-Watz, H. 1997. The Yersinia Yop virulon: a bacterial system for subverting eukaryotic cells. Mol. Microbiol. 23, 861–867.

    Article  CAS  PubMed  Google Scholar 

  • Covacci, A., Censini, S., Bugnoli, M., Petracca, R., Burroni, D., Macchia, G., Massone, A., Papini, E., Xiang, Z., and Figura, N. 1993. Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc. Natl. Acad. Sci. USA 90, 5791–5795.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cover, T.L. 1996. The vacuolating cytotoxin of Helicobacter pylori. Mol. Microbiol. 20, 241–246.

    Article  CAS  PubMed  Google Scholar 

  • 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 Res. 63, 951–957.

    CAS  PubMed  Google Scholar 

  • Delahay, R.M. and Frankel, G. 2002. Coiled-coil proteins associated with type III secretion systems: a versatile domain revisited. Mol. Microbiol. 45, 905–916.

    Article  CAS  PubMed  Google Scholar 

  • Dossumbekova, A., Prinz, C., Mages, J., Lang, R., Kusters, J.G., van Vliet, A.H.M., Reindl, W., Backert, S., Saur, D., Schmid, R.M., et al. 2006. Helicobacter pylori HopH (OipA) and bacterial pathogenicity: genetic and functional genomic analysis of hopH gene polymorphisms. J. Infect. Dis. 194, 1346–1355.

    Article  CAS  PubMed  Google Scholar 

  • Galmiche, A., Rassow, J., Doye, A., Cagnol, S., Chambard, J.C., Contamin, S., de Thillot, V., Just, I., Ricci, V., Solcia, E., et al. 2000. The N-terminal 34 kDa fragment of Helicobacter pylori vacuolating cytotoxin targets mitochondria and induces cytochrome c release. EMBO J. 19, 6361–6370.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Garufi, G., Butler, E., and Missiakas, D. 2008. ESAT-6-like protein secretion in Bacillus anthracis. J. Bacteriol. 190, 7004–7011.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gueroult, M., Picot, D., Abi,-Ghanem, J., Hartmann, B., and Baaden, M. 2010. How cations can assist DNase I in DNA binding and hydrolysis. PLoS Comput. Biol. 6, e1001000.

    Article  Google Scholar 

  • Haraga, A. and Miller, S.I. 2006. A Salmonella type III secretion effector interacts with the mammalian serine/threonine protein kinase PKN1. Cell. Microbiol. 8, 837–846.

    Article  CAS  PubMed  Google Scholar 

  • Hatakeyama, M. and Higashi, H. 2005. Helicobacter pylori CagA: a new paradigm for bacterial carcinogenesis. Cancer Sci. 96, 835–843.

    Article  CAS  PubMed  Google Scholar 

  • Hirano, T., Kobayashi, R., and Hirano, M. 1997. Condensins, chromosome condensation protein complexes containing XCAP-C, XCAP-E and a Xenopus homolog of the Drosophila Barren protein. Cell 89, 511–521.

    Article  CAS  PubMed  Google Scholar 

  • Hirano, T. 2002. The ABCs of SMC proteins: two-armed ATPases for chromosome condensation, cohesion, and repair. Gene Dev. 16, 399–414.

    Article  CAS  PubMed  Google Scholar 

  • Johnson, W.M. and Lior, H. 1998. A new heat-labile cytolethal distending toxin (CLDT) produced by Campylobacter spp. Microb. Pathog. 4, 115–126.

    Article  Google Scholar 

  • Lara-Tejero, M. and Galán, J.E. 2000. A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein. Science 290, 354–357.

    Article  CAS  PubMed  Google Scholar 

  • Lee, J.H., Jun, S.H., Baik, S.C., Kim, D.R., Park, J.Y., Lee, Y.S., Choi, C.H., and Lee, J.C. 2012. Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori. J. Microbiol. Methods 91, 490–496.

    Article  CAS  PubMed  Google Scholar 

  • Lorenzo, H.K., Susin, S.A., Penninger, J., and Kroemer, G. 1999. Apoptosis inducing factor (AIF): a phylogenetically old, caspase-independent effector of cell death. Cell Death Differ. 6, 516–524.

    Article  CAS  PubMed  Google Scholar 

  • Michaelis, C., Ciosk, R., and Nasmyth, K. 1997. Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell 91, 35–45.

    Article  CAS  PubMed  Google Scholar 

  • Negrini, R., Savio, A., Poiesi, C., Appelmelk, B.J., Buffoli, F., Paterlini, A., Cesari, P., Graffeo, M., Vaira, D., and Franzin, G. 1996. Antigenic mimicry between Helicobacter pylori and gastric mucosa in the pathogenesis of body atrophic gastritis. Gastroenterology 111, 655–665.

    Article  CAS  PubMed  Google Scholar 

  • Nomura, A., Stemmermann, G.N., Chyou, P.H., Kato, I., Perez-Perez, G.I., and Blaser, M.J. 1991. Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. N. Engl. J. Med. 325, 1132–1136.

    Article  CAS  PubMed  Google Scholar 

  • Okuda, J., Toyotome, T., Kataoka, N., Ohno, M., Abe, H., Shimura, Y., Seyedarabi, A., Pickersgill, R., and Sasakawa, C. 2005. Shigella effector IpaH9.8 binds to a splicing factor U2AF 35 to modulate host immune responses. Biochem. Biophys. Res. Commun. 333, 531–539.

    Article  CAS  PubMed  Google Scholar 

  • Parsonnet, J., Friedman, G.D., Vandersteen, D.P., Chang, Y., Vogelman, J.H., Orentreich, N., and Sibley, R.K. 1991. Helicobacter pylori infection and the risk of gastric carcinoma. N. Engl. J. Med. 325, 1127–1131.

    Article  CAS  PubMed  Google Scholar 

  • Pickett, C.L., Pesci, E.C., Cottle, D.L., Russell, G., Erdem, A.N., and Zeytin, H. 1996. Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campylobacter sp. cdtB gene. Infect. Immun. 64, 2070–2078.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Riddick, G. and Macara, I.G. 2005. A systems analysis of importin-α–β mediated nuclear protein import. J. Cell Biol. 168, 1027–1038.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Salvesen, G.S. and Dixit, V.M. 1999. Caspase activation: the induced-proximity model. Proc. Natl. Acad. Sci. USA 96, 10964–10967.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sharma, C.M., Hoffmann, S., Darfeuille, F., Reignier, J., Findeiß, S., Sittka, A., Chabas, S., Reiche, K., Hackermuller, J., Reinhardt, R., et al. 2010. The primary transcriptome of the major human pathogen Helicobacter pylori. Nature 464, 250–255.

    Article  CAS  PubMed  Google Scholar 

  • Thornberry, N.A. and Lazebnik, Y. 1998. Caspases: enemies within. Science 281, 1312–1316.

    Article  CAS  PubMed  Google Scholar 

  • Toller, I.M., Neelsen, K.J., Steger, M., Hartung, M.L., Hottiger, M.O., Stucki, M., Kalali, B., Gerhard, M., Sartori, A.A., Lopes, M., et al. 2011. Carcinogenic bacterial pathogen Helicobacter pylori triggers DNA double-strand breaks and a DNA damage response in its host cells. Proc. Natl. Acad. Sci. USA 108, 14944–14949.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Trigg, J., Gutwin, K., Keating, A.E., and Berger, B. 2011. Multicoil2: predicting coiled coils and their oligomerization states from sequence in the twilight zone. PLoS One 6, e23519.

    Article  Google Scholar 

  • Tsuji, S., Kawai, N., Tsujii, M., Kawano, S., and Hori, M. 2003. Review article: inflammation-related promotion of gastrointestinal carcinogenesis-a perigenetic pathway. Aliment Pharmacol. Ther. 18(S1), 82–89.

    Article  CAS  PubMed  Google Scholar 

  • Vincent, T.L., Green, P.J., and Woolfson, D.N. 2013. LOGICOILmulti-state prediction of coiled-coil oligomeric state. Bioinformatics 29, 69–76.

    Article  CAS  PubMed  Google Scholar 

  • Warren, J.R. and Marshall, B. 1983. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 321, 1273–1275.

    Article  Google Scholar 

  • Wroblewski, L.E., Peek, R.M., and Wilson, K.T. 2010. Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clinic. Microbiol. Rev. 23, 713–739.

    Article  CAS  Google Scholar 

  • Yachdav, G., Kloppmann, E., Kajan, L., Hecht, M., Goldberg, T., Hamp, T., Hönigschmid, P., Schafferhans, A., Roos, M., Bernhofer, M., et al. 2014. PredictProtein-an open resource for online prediction of protein structural and functional features. Nucleic Acids Res. 42, W337–W343.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yu, H.J., Lim, D., and Lee, H.S. 2003. Characterization of a novel single-stranded RNA mycovirus in Pleurotus ostreatus. Virology 314, 9–15.

    Article  CAS  PubMed  Google Scholar 

  • Zhang, D., Iyer, L.M., and Aravind, L. 2011. A novel immunity system for bacterial nucleic acid degrading toxins and its recruitment in various eukaryotic and DNA viral systems. Nucleic Acids Res. 39, 4532–4552.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, Gyeongsang National University School of Medicine, Jinju, 52727, Republic of Korea

    Young Chul Kwon, Myung-Je Cho, Woo-Kon Lee, Hyung-Lyun Kang, Jae-Young Song & Seung Chul Baik

  2. Division of Life Science and Research Institute of Life Sciences, Gyeongsang National University, Jinju, 52828, Republic of Korea

    Sinil Kim & Hyeon Su Ro

  3. Department of Life Science and Biotechnology, Soonchunhyang University, Asan, 31538, Republic of Korea

    Yong Seok Lee

  4. Department of Microbiology, Kyungpook National University School of Medicine, Daegu, 41944, Republic of Korea

    Je Chul Lee

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  1. Young Chul Kwon
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Correspondence to Seung Chul Baik or Hyeon Su Ro.

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Kwon, Y.C., Kim, S., Lee, Y.S. et al. Novel nuclear targeting coiled-coil protein of Helicobacter pylori showing Ca2+-independent, Mg2+-dependent DNase I activity. J Microbiol. 54, 387–395 (2016). https://doi.org/10.1007/s12275-016-5631-9

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  • DOI: https://doi.org/10.1007/s12275-016-5631-9

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