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Inactivation of the dnaK gene in Clostridium difficile 630 Delta erm yields a temperature-sensitive phenotype and increases biofilm-forming ability

Jain, Shailesh, Smyth, Deborah, O'Hagan, Barry, Heap, John, McMullan, Geoff, Minton, Nigel and Ternan, Nigel (2017) Inactivation of the dnaK gene in Clostridium difficile 630 Delta erm yields a temperature-sensitive phenotype and increases biofilm-forming ability. Scientific Reports, 7 . [Journal article]

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URL: http://rdcu.be/Bg7E

DOI: DOI:10.1038/s41598-017-17583-9

Abstract

Clostridium difficile infection is a growing problem in healthcare settings worldwide and resultsin a considerable socioeconomic impact. New hypervirulent strains and acquisition of antibioticresistance exacerbates pathogenesis; however, the survival strategy of C. difficile in the challenginggut environment still remains incompletely understood. We previously reported that clinically relevant heat-stress (37–41 °C) resulted in a classical heat-stress response with up-regulation of cellular chaperones. We used ClosTron to construct an insertional mutation in the dnaK gene of C. difficile 630 Δerm. The dnaK mutant exhibited temperature sensitivity, grew more slowly than C. difficile 630 Δerm and was less thermotolerant. Furthermore, the mutant was non-motile, had 4-fold lower expression of the fliC gene and lacked flagella on the cell surface. Mutant cells were some 50% longer than parental strain cells, and at optimal growth temperatures, they exhibited a 4-fold increase in the expression of class I chaperone genes including GroEL and GroES. Increased chaperone expression, in addition to the non-flagellated phenotype of the mutant, may account for the increased biofilm formation observed. Overall, the phenotype resulting from dnaK disruption is more akin to that observed in Escherichia coli dnaK mutants, rather than those in the Gram-positive model organism Bacillus subtilis.Clostridium difficile is recognised as the most common cause of infectious antibiotic-associated

Item Type:Journal article
Keywords:Clostridium difficile gut pathogen ClosTron, chaperone gene dnaK
Faculties and Schools:Faculty of Life and Health Sciences > School of Biomedical Sciences
Faculty of Life and Health Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute > Nutrition Innovation Centre for Food and Health (NICHE)
Biomedical Sciences Research Institute > Genomic Medicine
Biomedical Sciences Research Institute
ID Code:39185
Deposited By: Dr Nigel Ternan
Deposited On:19 Dec 2017 09:37
Last Modified:19 Dec 2017 09:37

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