Journal article
Dimethyl adenosine transferase (KsgA) contributes to cell-envelope fitness in Salmonella Enteritidis
Microbiological research, Vol.216, pp.108-119
11/2018
Handle:
https://hdl.handle.net/2376/103034
PMCID: PMC6628923
PMID: 30269850
Abstract
We previously reported that inactivation of a universally conserved dimethyl adenosine transferase (KsgA) attenuates virulence and increases sensitivity to oxidative and osmotic stress in Salmonella Enteritidis. Here, we show a role of KsgA in cell-envelope fitness as a potential mechanism underlying these phenotypes in Salmonella. We assessed structural integrity of the cell-envelope by transmission electron microscopy, permeability barrier function by determining intracellular accumulation of ethidium bromide and electrophysical properties by dielectrophoresis, an electrokinetic tool, in wild-type and ksgA knock-out mutants of S. Enteritidis. Deletion of ksgA resulted in disruption of the structural integrity, permeability barrier and distorted electrophysical properties of the cell-envelope. The cell-envelope fitness defects were alleviated by expression of wild-type KsgA (WT-ksgA) but not by its catalytically inactive form (ksgAE66A), suggesting that the dimethyl transferase activity of KsgA is important for cell-envelope fitness in S. Enteritidis. Upon expression of WT-ksgA and ksgAE66A in inherently permeable E. coli cells, the former strengthened and the latter weakened the permeability barrier, suggesting that KsgA also contributes to the cell-envelope fitness in E. coli. Lastly, expression of ksgAE66A exacerbated the cell-envelope fitness defects, resulting in impaired S. Enteritidis interactions with human intestinal epithelial cells, and human and avian phagocytes. This study shows that KsgA contributes to cell-envelope fitness and opens new avenues to modulate cell-envelopes via use of KsgA-antagonists.
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Details
- Title
- Dimethyl adenosine transferase (KsgA) contributes to cell-envelope fitness in Salmonella Enteritidis
- Creators
- Kim Lam Chiok - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USANarayan C Paul - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAEzekiel O Adekanmbi - Department of Chemical and Materials Engineering, University of Idaho, Moscow, Idaho 83844-1021, USASoumya K Srivastava - Department of Chemical and Materials Engineering, University of Idaho, Moscow, Idaho 83844-1021, USADevendra H Shah - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA
- Publication Details
- Microbiological research, Vol.216, pp.108-119
- Academic Unit
- Veterinary Microbiology and Pathology, Department of
- Publisher
- Elsevier GmbH
- Identifiers
- 99900546542201842
- Language
- English
- Resource Type
- Journal article