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https://hdl.handle.net/2440/76193
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Type: | Journal article |
Title: | Role of lipid rafts and flagellin in invasion of colonic epithelial cells by Shiga-toxigenic Escherichia coli O113:H21 |
Author: | Rogers, T. Thorpe, C. Paton, A. Paton, J. |
Citation: | Infection and Immunity, 2012; 80(8):2858-2867 |
Publisher: | Amer Soc Microbiology |
Issue Date: | 2012 |
ISSN: | 0019-9567 1098-5522 |
Editor: | McCormick, B.A. |
Statement of Responsibility: | Trisha J. Rogers, Cheleste M. Thorpe, Adrienne W. Paton, and James C. Paton |
Abstract: | Shiga-toxigenic Escherichia coli (STEC) O113:H21 strains that lack the locus of enterocyte effacement (LEE) efficiently invade eukaryotic cells in vitro, unlike LEE-positive O157:H7 strains. We used a fliC deletion mutant of the O113:H21 STEC strain 98NK2 (98NK2ΔfliC) to show that invasion of colonic epithelial (HCT-8) cells is heavily dependent on production of flagellin, even though adherence to the cells was actually enhanced in the mutant. Flagellin binds and signals through Toll-like receptor 5 (TLR5), but there was no evidence that either TLR5, the adaptor protein myeloid differentiation primary response gene 88 (MyD88), or the serine kinase interleukin-1 receptor-associated kinase (IRAK) were required for invasion of HCT-8 cells by strain 98NK2, as judged by transfection, RNA knockdown, or inhibitor studies. However, pretreatment of cells with anti-asialo-GM1 significantly decreased 98NK2 invasion (by 40.8%), while neuraminidase treatment (which cleaves terminal sialic acid residues, thus converting GM1 into asialo-GM1) significantly increased invasion (by 70.7%). Pretreatment of HCT-8 cells with either the cholesterol-depleting agent methyl-β-cyclodextrin (MβCD) or the tyrosine kinase inhibitor genistein significantly decreased invasion by 98NK2, indicating a potential role for lipid rafts in the invasion mechanism. Confocal microscopy also showed invading 98NK2 colocalized with lipid raft markers caveolin-1 and GM1. Interestingly, anti-asialo-GM1, neuraminidase, MβCD, and genistein have similar effects on the vestigial level of STEC invasion seen for STEC strain 98NK2ΔfliC, indicating that lipid rafts mediate a common step in flagellin-dependent and flagellin-independent cellular invasion. |
Keywords: | Colon Cell Line, Tumor Membrane Microdomains Epithelial Cells Humans G(M1) Ganglioside Flagellin RNA, Small Interfering Gene Expression Regulation, Bacterial RNA Interference Protein Binding Toll-Like Receptor 5 Shiga-Toxigenic Escherichia coli |
Rights: | Copyright © 2012, American Society for Microbiology. All Rights Reserved. |
DOI: | 10.1128/IAI.00336-12 |
Published version: | http://dx.doi.org/10.1128/iai.00336-12 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
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