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Cold Shock Response in Sporulating Bacillus subtilis and Its Effect on Spore Heat Resistance

Division of Food Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Sutton Bonington, Loughborough, Leicestershire LE12 5RD, United Kingdom

Received 25 February 2002/ Accepted 26 June 2002

Cold shock and ethanol and puromycin stress responses in sporulating Bacillus subtilis cells have been investigated. We show that a total of 13 proteins are strongly induced after a short cold shock treatment of sporulating cells. The cold shock pretreatment affected the heat resistance of the spores formed subsequently, with spores heat killed at 85 or 90°C being more heat resistant than the control spores while they were more heat sensitive than controls that were heat treated at 95 or 100°C. However, B. subtilis spores with mutations in the main cold shock proteins, CspB, -C, and -D, did not display decreased heat resistance compared to controls, indicating that these proteins are not directly responsible for the increased heat resistance of the spores. The disappearance of the stress proteins later in sporulation suggests that they cannot be involved in repairing heat damage during spore germination and outgrowth but must alter spore structure in a way which increases or decreases heat resistance. Since heat, ethanol, and puromycin stress produce similar proteins and similar changes in spore heat resistance while cold shock is different in both respects, these alterations appear to be very specific.