Bacteroides fragilis toxin rearranges the actin cytoskeleton of HT29/C1 cells without direct proteolysis of actin or decrease in F-actin content

Roxan F. Saidi, Kristin Jaeger, Marshall H. Montrose, Shaoguang Wu, Cynthia L. Sears

Research output: Contribution to journalArticlepeer-review

Abstract

Enterotoxigenic strains of B. fragilis associated with childhood diarrhea produce a 20 kD zinc metalloprotease toxin (BFT). BFT is reported to cleave G-actin in vitro and also causes dramatic rounding and rearrangement of the F-actin cytoskeleton in human intestinal epithelial cell lines (HT29 and HT29/C1). To test the hypothesis that the proteolysis of cellular actin by BFT in vivo may contribute to these alterations in morphology and cytoskeletal architecture, we assessed the F-actin content and the arrangement of the F- and G-actin cytoskeleton in BFT-treated HT29/C1 cells by spectrofluorimetry, confocal microscopy, and immunoblotting. BFT-treated cells were compared to cells treated with C. difficile toxin A (CDA) or cytochalasin D. Using spectrofluorimetric quantification, the F-actin content of BFT and cytochalasin D-treated cells was unchanged in contrast to a significant decrease in CDA-treated cells. By confocal microscopy, the arrangement of F and G-actin in all treated cells was markedly different than control cells. There was no change in the immunoblotting pattern of actin in the Triton-soluble or -insoluble cellular fractions of BFT-treated HT29/C1 cells. We conclude that BFT alters the F- and G-actin cytoskeletal architecture of HT29/C1 cells without direct proteolysis of actin or decrease in F-actin content.

Original languageEnglish (US)
Pages (from-to)159-165
Number of pages7
JournalCell Motility and the Cytoskeleton
Volume37
Issue number2
DOIs
StatePublished - Jun 26 1997

Keywords

  • Bacteroides fragilis
  • Enterotoxigenic Bacteroides fragilis
  • Intestinal cells
  • Toxin
  • actin
  • cytoskeleton

ASJC Scopus subject areas

  • Structural Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Bacteroides fragilis toxin rearranges the actin cytoskeleton of HT29/C1 cells without direct proteolysis of actin or decrease in F-actin content'. Together they form a unique fingerprint.

Cite this