The protease CspB is essential for initiation of cortex hydrolysis and dipicolinic acid (DPA) release during germination of spores of type A food poisoning isolates Free

Abstract

The genome of the food poisoning isolate SM101 encodes a subtilisin-like protease, CspB, upstream of the gene encoding the enzyme essential for degradation of the peptidoglycan cortex during spore germination. SleC is an inactive pro-SleC in dormant spores that is converted to active SleC during spore germination and Csp proteases convert pro-SleC to the active enzyme . In this work, the germination and viability of spores of a deletion mutant of strain SM101, as well as expression, were studied. The gene was expressed only during sporulation, and only in the mother cell compartment. spores were unable to germinate significantly with either a rich nutrient medium, KCl, or a 1 : 1 chelate of Ca and dipicolinic acid (DPA); the viability of these spores was ∼10-fold lower than that of wild-type spores, although and wild-type spores had similar viability on plates containing lysozyme, and spores could not process inactive pro-SleC into active SleC during spore germination. Germination of spores was blocked prior to DPA release and cortex hydrolysis, and germination and viability defects in these spores were complemented by an ectopic . These results indicate that Csp proteases are essential to generate active SleC and allow cortex hydrolysis early in spore germination. However, Csp proteases likely play another role in spore germination, since spores did not release DPA upon exposure to germinants, while spores have been shown previously to release DPA, albeit slowly, upon exposure to germinants.

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2009-10-01
2024-03-29
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