Mechanical Fractionation Reveals Structural Requirements for Enteropathogenic Escherichia coli Tir Insertion into Host Membranes
Open Access
- 1 July 2000
- journal article
- research article
- Published by American Society for Microbiology in Infection and Immunity
- Vol. 68 (7) , 4344-4348
- https://doi.org/10.1128/iai.68.7.4344-4348.2000
Abstract
Enteropathogenic Escherichia coli (EPEC) inserts its receptor for intimate adherence (Tir) into host cell membranes by using a type III secretion system. Detergents are frequently used to fractionate infected host cells to investigate bacterial protein delivery into mammalian cells. In this study, we found that the Triton X-100-soluble membrane fraction from EPEC-infected HeLa cells was contaminated with bacterial proteins. We therefore applied a mechanical method of cell lysis and ultracentrifugation to fractionate infected HeLa cells to investigate the biology and biochemistry of Tir delivery and translocation. This method demonstrates that the translocation of Tir into the host cell membrane requires its transmembrane domains, but not tyrosine phosphorylation or binding to Tir's ligand, intimin.Keywords
This publication has 26 references indexed in Scilit:
- Protein translocation into mitochondria: the role of TIM complexesTrends in Cell Biology, 2000
- Enteropathogenic Escherichia coli translocated intimin receptor, Tir, requires a specific chaperone for stable secretionMolecular Microbiology, 1999
- Identification of the intimin-binding domain of Tir of enteropathogenic Escherichia coliCellular Microbiology, 1999
- Protein tyrosine kinases in bacterial pathogens are associated with virulence and production of exopolysaccharideThe EMBO Journal, 1999
- Binding of intimin from enteropathogenic Escherichia coli to Tir and to host cellsMolecular Microbiology, 1999
- Initial binding of Shiga toxin‐producing Escherichia coli to host cells and subsequent induction of actin rearrangements depend on filamentous EspA‐containing surface appendagesMolecular Microbiology, 1998
- A novel EspA-associated surface organelle of enteropathogenic Escherichia coli involved in protein translocation into epithelial cellsThe EMBO Journal, 1998
- Intracellular targeting of exoenzyme S of Pseudomonas aeruginosa via type III‐dependent translocation induces phagocytosis resistance, cytotoxicity and disruption of actin microfilamentsMolecular Microbiology, 1997
- Cell‐surface‐bound Yersinia translocate the protein tyrosine phosphatase YopH by a polarized mechanism into the target cellMolecular Microbiology, 1995
- Role of the Escherichia coli DnaK and DnaJ heat shock proteins in the initiation of bacteriophage lambda DNA replication.Proceedings of the National Academy of Sciences, 1988