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Journal of Bacteriology, April 2009, p. 2711-2720, Vol. 191, No. 8
0021-9193/09/$08.00+0     doi:10.1128/JB.01832-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

SleC Is Essential for Cortex Peptidoglycan Hydrolysis during Germination of Spores of the Pathogenic Bacterium Clostridium perfringens{triangledown} ,{dagger}

Daniel Paredes-Sabja,1 Peter Setlow,3 and Mahfuzur R. Sarker1,2*

Department of Biomedical Sciences, College of Veterinary Medicine,1 Department of Microbiology, College of Science, Oregon State University, Corvallis, Oregon 97331,2 Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 060303

Received 29 December 2008/ Accepted 28 January 2009

Clostridial spore germination requires degradation of the spore's peptidoglycan (PG) cortex by cortex-lytic enzymes (CLEs), and two Clostridium perfringens CLEs, SleC and SleM, degrade cortex PG in vitro. We now find that only SleC is essential for cortex hydrolysis and viability of C. perfringens spores. C. perfringens sleC spores did not germinate completely with nutrients, KCl, or a 1:1 chelate of Ca2+ and dipicolinic acid (Ca-DPA), and the colony-forming efficiency of sleC spores was 103-fold lower than that of wild-type spores. However, sleC spores incubated with various germinants released most of their DPA, although slower than wild-type or sleM spores, and DPA release from sleC sleM spores was very slow. In contrast, germination and viability of sleM spores were similar to that of wild-type spores, although sleC sleM spores had 105-fold-lower viability. These results allow the following conclusions about C. perfringens spore germination: (i) SleC is essential for cortex hydrolysis; (ii) although SleM can degrade cortex PG in vitro, this enzyme is not essential; (iii) action of SleC alone or with SleM can accelerate DPA release; and (iv) Ca-DPA does not trigger spore germination by activation of CLEs.

* Corresponding author. Mailing address: Department of Biomedical Sciences, Oregon State University, 216 Dryden Hall, Corvallis, OR 97331. Phone: (541) 737-6918. Fax: (541) 737-2730. E-mail: sarkerm{at}oregonstate.edu

{triangledown} Published ahead of print on 13 February 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, April 2009, p. 2711-2720, Vol. 191, No. 8
0021-9193/09/$08.00+0     doi:10.1128/JB.01832-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Paredes-Sabja, D., Udompijitkul, P., Sarker, M. R. (2009). Inorganic Phosphate and Sodium Ions Are Cogerminants for Spores of Clostridium perfringens Type A Food Poisoning-Related Isolates. Appl. Environ. Microbiol. 75: 6299-6305 [Abstract] [Full Text]  
  • Paredes-Sabja, D., Setlow, P., Sarker, M. R. (2009). The protease CspB is essential for initiation of cortex hydrolysis and dipicolinic acid (DPA) release during germination of spores of Clostridium perfringens type A food poisoning isolates. Microbiology 155: 3464-3472 [Abstract] [Full Text]  
  • Paredes-Sabja, D., Setlow, P., Sarker, M. R. (2009). GerO, a Putative Na+/H+-K+ Antiporter, Is Essential for Normal Germination of Spores of the Pathogenic Bacterium Clostridium perfringens. J. Bacteriol. 191: 3822-3831 [Abstract] [Full Text]  
  • Paredes-Sabja, D., Setlow, P., Sarker, M. R. (2009). Role of GerKB in Germination and Outgrowth of Clostridium perfringens Spores. Appl. Environ. Microbiol. 75: 3813-3817 [Abstract] [Full Text]  


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