PMID:21367479

Citation	Söderholm, H, Lindström, M, Somervuo, P, Heap, J, Minton, N, Lindén, J and Korkeala, H (2011) cspB encodes a major cold shock protein in Clostridium botulinum ATCC 3502. Int. J. Food Microbiol. 146:23-30
Abstract	The relative expression of three cold shock protein coding genes (cspA, cspB and cspC) of Clostridium botulinum ATCC 3502 was studied with quantitative RT-PCR analysis following a cold shock shift from 37 °C to 15 °C. A significant increase in the relative expression of all three genes was observed upon the temperature downshift. To validate these findings, single-gene insertional inactivation of cspA, cspB and cspC was undertaken with the ClosTron gene knock-out system. In growth experiments, mutations in cspB or cspC, but not cspA, resulted in a cold-sensitive phenotype. No growth of the cspB mutant was observed at 15°C over a ten day period, whereas at 20 °C the growth rate was 70% lower than that of wild type strain. The growth rate of cspC mutant was 70% and 80% lower than the growth rate of the wild type strain at 15 °C and 20 °C, respectively. At 37 °C the growth of cspB mutant did not differ from, but the growth rate of cspC mutant was 30% lower than, that of the wild type strain. The cspA mutant grew somewhat faster than the wild type strain at all studied temperatures. Since the inactivation of cspB resulted in the most prominent defect in growth at low temperatures, we suggest that cspB encodes the major cold shock protein of C. botulinum ATCC 3502. Understanding the mechanisms behind cold tolerance of C. botulinum helps to evaluate the safety risks this foodborne pathogen poses in the modern food industry.
Links	PubMed Online version:10.1016/j.ijfoodmicro.2011.01.033
Keywords	Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; Clostridium botulinum/genetics; Clostridium botulinum/growth & development; Clostridium botulinum/metabolism; Cold Shock Proteins and Peptides/genetics; Cold Shock Proteins and Peptides/metabolism; Cold Temperature; DNA, Bacterial/genetics; Gene Expression Regulation, Bacterial; Gene Knockout Techniques; Molecular Sequence Data; Mutation; Sequence Analysis, DNA
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
CLOBH:A5I1L8		GO:0070417: cellular response to cold	ECO:0000315:		P		table 4. the growth rate of the cspB mutant is significantly lower than wild type at 15 and 20 degrees C.	complete CACAO 4765
CLOBH:A5I1L8	involved_in	GO:0070417: cellular response to cold	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

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