Enteropathogenic Escherichia coli effector EspF interacts with host protein Abcf2

Cell Microbiol. 2007 Mar;9(3):680-93. doi: 10.1111/j.1462-5822.2006.00820.x. Epub 2006 Oct 25.

Abstract

Enteropathogenic Escherichia coli (EPEC) is a major causative agent of infant diarrhoea in developing countries. The EspF effector protein is injected from EPEC into host cells via a type III secretion system and is involved in the disruption of host intestinal barrier function. In addition, EspF is sorted to mitochondria and has a role in initiating the mitochondrial death pathway. To clarify the manner in which EspF affects host cells, we sought to identify eukaryotic EspF-binding proteins using affinity purification. Abcf2, a protein of unknown function and member of the ABC-transporter family, bound EspF in this assay. An interaction between EspF and Abcf2 was confirmed in a yeast two-hybrid system, by colocalization and by co-immunoprecipitation from EPEC-infected cells. Levels of Abcf2 were decreased in cells infected with EPEC in an EspF dose-dependent manner. Knock-down of Abcf2 expression by RNA interference increased EspF-induced caspase 9 and caspase 3 cleavage. In addition, Abcf2-knocked down cells showed increased caspase 3 cleavage upon treatment with the apoptosis inducing agent staurosporine. These results indicate that EspF induces or facilitates host cell death by targeting and interfering with the putative protective function of Abcf2.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / metabolism*
  • Amino Acid Sequence
  • Blotting, Western
  • Caco-2 Cells
  • Carrier Proteins / metabolism*
  • Escherichia coli Proteins / metabolism*
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins
  • Microscopy, Fluorescence
  • Mutation
  • Protein Binding
  • RNA, Small Interfering / genetics
  • Two-Hybrid System Techniques

Substances

  • ABCF2 protein, human
  • ATP-Binding Cassette Transporters
  • Carrier Proteins
  • Escherichia coli Proteins
  • EspFU protein, E coli
  • Intracellular Signaling Peptides and Proteins
  • RNA, Small Interfering