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Volume 157, Issue 4

The muramidase EtgA from enteropathogenic is required for efficient type III secretion Free

Abstract

Enteropathogenic (EPEC) is an important cause of infectious diarrhoea. It colonizes human intestinal epithelial cells by delivering effector proteins into the host cell cytoplasm via a type III secretion system (T3SS) encoded within the chromosomal locus of enterocyte effacement (LEE). The LEE pathogenicity island also encodes a lytic transglycosylase (LT) homologue named EtgA. In the present work we investigated the significance of EtgA function in type III secretion (T3S). Purified recombinant EtgA was found to have peptidoglycan lytic activity . Consistent with this function, signal peptide processing and bacterial cell fractionation revealed that EtgA is a periplasmic protein. EtgA possesses the conserved glutamate characteristic of the LT family, and we show here that it is essential for enzymic activity. Overproduction of EtgA in EPEC inhibits bacterial growth and induces cell lysis unless the predicted catalytic glutamate is mutated. An mutant is attenuated for T3S, red blood cell haemolysis and EspA filamentation. BfpH, a plasmid-encoded putative LT, was not able to functionally replace EtgA. Overall, our results indicate that the muramidase activity of EtgA is not critical but makes a significant contribution to the efficiency of the T3S process.

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Keyword(s): A/E, attaching and effacing , BDP, E. coli BL21 DE3 pLysS , BFP, bundle-forming pilus , CBB, Coomassie brilliant blue , EAF, EPEC adherence factor , EHEC, enterohaemorrhagic Escherichia coli , EPEC, enteropathogenic Escherichia coli , LEE, locus of enterocyte effacement , LT, lytic transglycosylase , MBP, maltose binding protein , PG, peptidoglycan , RBC, red blood cell , T3S, type III secretion , T3SS, type III secretion system and T4SS, type IV secretion system
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2011-04-01
2024-04-23
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The muramidase EtgA from enteropathogenic Escherichia coli is required for efficient type III secretion
Microbiology 157, 1145 (2011); https://doi.org/10.1099/mic.0.045617-0
/content/journal/micro/10.1099/mic.0.045617-0
/content/journal/micro/10.1099/mic.0.045617-0
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