1887

Abstract

is the most common cause of enteric disease and presents a major burden on healthcare systems globally due in part to the observed rapid rise in antibiotic resistance. The ability of to form endospores is a key feature in the organism’s pathogenesis and transmission, and contributes greatly to its resilient nature. Endospores are highly resistant to disinfection, allowing them to persist on hospital surfaces. In order for the organism to cause disease, the spores must germinate and revert to a vegetative form. While spore germination in spp. is well understood, very little is known about this process in . Here we report the characterization of SleC (CD0551) from 630. Bioinformatic analysis of SleC indicated a multi-domained protein possessing a peptidoglycan-binding (PGB) domain, a SpoIID/LytB domain and an undefined N-terminal region. We have confirmed that SleC is an exo-acting lytic transglycosylase with the catalytic activity localized to the N-terminal region. Additionally, we have shown that both the N-terminal catalytic domain and the C-terminal PGB domain require muramyl-δ-lactam for substrate binding. As with carbohydrate-binding modules from cellulases and xylanases, the PGB domain may be responsible for increasing the processivity of SleC by concentrating the enzyme at the surface of the substrate.

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2014-01-01
2019-12-05
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References

  1. Adams C. M., Eckenroth B. E., Putnam E. E., Doublié S., Shen A.. ( 2013;). Structural and functional analysis of the CspB protease required for Clostridium spore germination. . PLoS Pathog 9:, e1003165. [CrossRef][PubMed]
    [Google Scholar]
  2. Ali M. K., Hayashi H., Karita S., Goto M., Kimura T., Sakka K., Ohmiya K.. ( 2001;). Importance of the carbohydrate-binding module of Clostridium stercorarium Xyn10B to xylan hydrolysis. . Biosci Biotechnol Biochem 65:, 41–47. [CrossRef][PubMed]
    [Google Scholar]
  3. Bolam D. N., Ciruela A., McQueen-Mason S., Simpson P., Williamson M. P., Rixon J. E., Boraston A., Hazlewood G. P., Gilbert H. J.. ( 1998;). Pseudomonas cellulose-binding domains mediate their effects by increasing enzyme substrate proximity. . Biochem J 331:, 775–781.[PubMed]
    [Google Scholar]
  4. Boraston A. B., Kwan E., Chiu P., Warren R. A., Kilburn D. G.. ( 2003;). Recognition and hydrolysis of noncrystalline cellulose. . J Biol Chem 278:, 6120–6127. [CrossRef][PubMed]
    [Google Scholar]
  5. Boraston A. B., Bolam D. N., Gilbert H. J., Davies G. J.. ( 2004;). Carbohydrate-binding modules: fine-tuning polysaccharide recognition. . Biochem J 382:, 769–781. [CrossRef][PubMed]
    [Google Scholar]
  6. Burns D. A., Heap J. T., Minton N. P.. ( 2010a;). SleC is essential for germination of Clostridium difficile spores in nutrient-rich medium supplemented with the bile salt taurocholate. . J Bacteriol 192:, 657–664. [CrossRef][PubMed]
    [Google Scholar]
  7. Burns D. A., Heap J. T., Minton N. P.. ( 2010b;). Clostridium difficile spore germination: an update. . Res Microbiol 161:, 730–734. [CrossRef][PubMed]
    [Google Scholar]
  8. Callewaert L., Walmagh M., Michiels C. W., Lavigne R.. ( 2011;). Food applications of bacterial cell wall hydrolases. . Curr Opin Biotechnol 22:, 164–171. [CrossRef][PubMed]
    [Google Scholar]
  9. Carroll K. C., Bartlett J. G.. ( 2011;). Biology of Clostridium difficile: implications for epidemiology and diagnosis. . Annu Rev Microbiol 65:, 501–521. [CrossRef][PubMed]
    [Google Scholar]
  10. Chalut C., Charpentier X., Remy M.-H., Masson J.-M.. ( 2001;). Differential responses of Escherichia coli cells expressing cytoplasmic domain mutants of penicillin-binding protein 1b after impairment of penicillin-binding proteins 1a and 3. . J Bacteriol 183:, 200–206. [CrossRef][PubMed]
    [Google Scholar]
  11. Courtin P., Miranda G., Guillot A., Wessner F., Mézange C., Domakova E., Kulakauskas S., Chapot-Chartier M. P.. ( 2006;). Peptidoglycan structure analysis of Lactococcus lactis reveals the presence of an l,d-carboxypeptidase involved in peptidoglycan maturation. . J Bacteriol 188:, 5293–5298. [CrossRef][PubMed]
    [Google Scholar]
  12. Deakin L. J., Clare S., Fagan R. P., Dawson L. F., Pickard D. J., West M. R., Wren B. W., Fairweather N. F., Dougan G., Lawley T. D.. ( 2012;). The Clostridium difficile spo0A gene is a persistence and transmission factor. . Infect Immun 80:, 2704–2711. [CrossRef][PubMed]
    [Google Scholar]
  13. Dideberg O., Charlier P., Dive G., Joris B., Frère J. M., Ghuysen J. M.. ( 1982;). Structure of a Zn2+-containing d-alanyl-d-alanine-cleaving carboxypeptidase at 2.5 Å resolution. . Nature 299:, 469–470. [CrossRef][PubMed]
    [Google Scholar]
  14. Dowd M. M., Orsburn B., Popham D. L.. ( 2008;). Cortex peptidoglycan lytic activity in germinating Bacillus anthracis spores. . J Bacteriol 190:, 4541–4548. [CrossRef][PubMed]
    [Google Scholar]
  15. Foster S. J.. ( 1991;). Cloning, expression, sequence analysis and biochemical characterization of an autolytic amidase of Bacillus subtilis 168 trpC2. . J Gen Microbiol 137:, 1987–1998. [CrossRef][PubMed]
    [Google Scholar]
  16. Ghantoji S. S., Sail K., Lairson D. R., DuPont H. L., Garey K. W.. ( 2010;). Economic healthcare costs of Clostridium difficile infection: a systematic review. . J Hosp Infect 74:, 309–318. [CrossRef][PubMed]
    [Google Scholar]
  17. Heffron J. D., Sherry N., Popham D. L.. ( 2011;). In vitro studies of peptidoglycan binding and hydrolysis by the Bacillus anthracis germination-specific lytic enzyme SleB. . J Bacteriol 193:, 125–131. [CrossRef][PubMed]
    [Google Scholar]
  18. Hizukuri Y., Morton J. F., Yakushi T., Kojima S., Homma M.. ( 2009;). The peptidoglycan-binding (PGB) domain of the Escherichia coli Pal protein can also function as the PGB domain in E. coli flagellar motor protein MotB. . J Biochem 146:, 219–229. [CrossRef][PubMed]
    [Google Scholar]
  19. Ishikawa S., Yamane K., Sekiguchi J.. ( 1998;). Regulation and characterization of a newly deduced cell wall hydrolase gene (cwlJ) which affects germination of Bacillus subtilis spores. . J Bacteriol 180:, 1375–1380.[PubMed]
    [Google Scholar]
  20. Jehl M.-A., Arnold R., Rattei T.. ( 2011;). Effective – a database of predicted secreted bacterial proteins. . Nucleic Acids Res 39: (Database issue), D591–D595. [CrossRef][PubMed]
    [Google Scholar]
  21. Jing X., Robinson H. R., Heffron J. D., Popham D. L., Schubot F. D.. ( 2012;). The catalytic domain of the germination-specific lytic transglycosylase SleB from Bacillus anthracis displays a unique active site topology. . Proteins 80:, 2469–2475. [CrossRef][PubMed]
    [Google Scholar]
  22. Kee V. R.. ( 2012;). Clostridium difficile infection in older adults: a review and update on its management. . Am J Geriatr Pharmacother 10:, 14–24. [CrossRef][PubMed]
    [Google Scholar]
  23. Kelly C. P., LaMont J. T.. ( 2008;). Clostridium difficile – more difficult than ever. . N Engl J Med 359:, 1932–1940. [CrossRef][PubMed]
    [Google Scholar]
  24. Kopp U., Roos M., Wecke J., Labischinski H.. ( 1996;). Staphylococcal peptidoglycan interpeptide bridge biosynthesis: a novel antistaphylococcal target. ? Microb Drug Resist 2:, 29–41. [CrossRef][PubMed]
    [Google Scholar]
  25. Kumazawa T., Masayama A., Fukuoka S., Makino S., Yoshimura T., Moriyama R.. ( 2007;). Mode of action of a germination-specific cortex-lytic enzyme, SleC, of Clostridium perfringens S40. . Biosci Biotechnol Biochem 71:, 884–892. [CrossRef][PubMed]
    [Google Scholar]
  26. Lawley T. D., Clare S., Deakin L. J., Goulding D., Yen J. L., Raisen C., Brandt C., Lovell J., Cooke F.. & other authors ( 2010;). Use of purified Clostridium difficile spores to facilitate evaluation of health care disinfection regimens. . Appl Environ Microbiol 76:, 6895–6900. [CrossRef][PubMed]
    [Google Scholar]
  27. Leffler D. A., Lamont J. T.. ( 2009;). Treatment of Clostridium difficile-associated disease. . Gastroenterology 136:, 1899–1912. [CrossRef][PubMed]
    [Google Scholar]
  28. Leggett M. J., McDonnell G., Denyer S. P., Setlow P., Maillard J. Y.. ( 2012;). Bacterial spore structures and their protective role in biocide resistance. . J Appl Microbiol 113:, 485–498. [CrossRef][PubMed]
    [Google Scholar]
  29. Maroo S., Lamont J. T.. ( 2006;). Recurrent Clostridium difficile.. Gastroenterology 130:, 1311–1316. [CrossRef][PubMed]
    [Google Scholar]
  30. Paredes-Sabja D., Sarker M. R.. ( 2011;). Germination response of spores of the pathogenic bacterium Clostridium perfringens and Clostridium difficile to cultured human epithelial cells. . Anaerobe 17:, 78–84. [CrossRef][PubMed]
    [Google Scholar]
  31. Paredes-Sabja D., Setlow P., Sarker M. R.. ( 2009;). SleC is essential for cortex peptidoglycan hydrolysis during germination of spores of the pathogenic bacterium Clostridium perfringens.. J Bacteriol 191:, 2711–2720. [CrossRef][PubMed]
    [Google Scholar]
  32. Pépin J., Valiquette L., Cossette B.. ( 2005;). Mortality attributable to nosocomial Clostridium difficile-associated disease during an epidemic caused by a hypervirulent strain in Quebec. . CMAJ 173:, 1037–1042. [CrossRef][PubMed]
    [Google Scholar]
  33. Popham D. L., Helin J., Costello C. E., Setlow P.. ( 1996;). Muramic lactam in peptidoglycan of Bacillus subtilis spores is required for spore outgrowth but not for spore dehydration or heat resistance. . Proc Natl Acad Sci U S A 93:, 15405–15410. [CrossRef][PubMed]
    [Google Scholar]
  34. Reid C. W., Blackburn N. T., Clarke A. J.. ( 2006;). Role of arginine residues in the active site of the membrane-bound lytic transglycosylase B from Pseudomonas aeruginosa.. Biochemistry 45:, 2129–2138. [CrossRef][PubMed]
    [Google Scholar]
  35. Rupnik M., Wilcox M. H., Gerding D. N.. ( 2009;). Clostridium difficile infection: new developments in epidemiology and pathogenesis. . Nat Rev Microbiol 7:, 526–536. [CrossRef][PubMed]
    [Google Scholar]
  36. Sarker M. R., Paredes-Sabja D.. ( 2012;). Molecular basis of early stages of Clostridium difficile infection: germination and colonization. . Future Microbiol 7:, 933–943. [CrossRef][PubMed]
    [Google Scholar]
  37. Setlow P.. ( 2003;). Spore germination. . Curr Opin Microbiol 6:, 550–556. [CrossRef][PubMed]
    [Google Scholar]
  38. Shimamoto S., Moriyama R., Sugimoto K., Miyata S., Makino S.. ( 2001;). Partial characterization of an enzyme fraction with protease activity which converts the spore peptidoglycan hydrolase (SleC) precursor to an active enzyme during germination of Clostridium perfringens S40 spores and analysis of a gene cluster involved in the activity. . J Bacteriol 183:, 3742–3751. [CrossRef][PubMed]
    [Google Scholar]
  39. Tillotson G. S., Tillotson J.. ( 2011;). Clostridium difficile – a moving target. . F1000 Med Rep 3:, 6. [CrossRef][PubMed]
    [Google Scholar]
  40. Tomme P., Van Tilbeurgh H., Pettersson G., Van Damme J., Vandekerckhove J., Knowles J., Teeri T., Claeyssens M.. ( 1988;). Studies of the cellulolytic system of Trichoderma reesei QM 9414. Analysis of domain function in two cellobiohydrolases by limited proteolysis. . Eur J Biochem 170:, 575–581. [CrossRef][PubMed]
    [Google Scholar]
  41. Ursinus A., van den Ent F., Brechtel S., de Pedro M., Höltje J. V., Löwe J., Vollmer W.. ( 2004;). Murein (peptidoglycan) binding property of the essential cell division protein FtsN from Escherichia coli.. J Bacteriol 186:, 6728–6737. [CrossRef][PubMed]
    [Google Scholar]
  42. Walk S. T., Micic D., Galecki A. T., Young V. B., Aronoff D. M.. ( 2013;). Understanding increased mortality in Clostridium difficile-infected older adults. . Clin Infect Dis 57:, 625–626. [CrossRef][PubMed]
    [Google Scholar]
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