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Efficient endosome-to-Golgi transport of Shiga toxin is dependent on dynamin and clathrin

Silje U Lauvrak1, Maria L Torgersen, Kirsten Sandvig

  • 1Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, 0310 Oslo, Norway.

Journal of Cell Science
|May 6, 2004
PubMed

Insights

Shiga toxin transport from endosomes to the Golgi requires dynamin (a protein involved in vesicle formation). Butyric acid treatment makes cells more sensitive to Shiga toxin and necessitates clathrin (a protein coating vesicles) for this transport.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Toxicology

Background:

  • Shiga toxin enters cells via endocytosis, potentially through clathrin-dependent or independent pathways.
  • Following internalization, the toxin undergoes retrograde transport to the Golgi and endoplasmic reticulum, ultimately reaching the cytosol to inhibit protein synthesis.
  • Previous studies indicate Shiga toxin uptake and retrograde transport can be influenced by factors like clathrin and dynamin.

Purpose of the Study:

  • To investigate the roles of dynamin and clathrin in the endosome-to-Golgi transport of Shiga toxin.
  • To determine if butyric acid, a known sensitizer to Shiga toxin, alters the dependence of toxin transport on dynamin and clathrin.
  • To elucidate the mechanisms by which Shiga toxin reaches the cytosol to exert its toxic effects.

Main Methods:

  • Utilized HeLa dyn(K44A) and BHK antisense clathrin heavy chain (CHC) cells for inducible expression of mutant dynamin and CHC antisense RNA, respectively.
  • Assessed Shiga toxin internalization in cells with blocked dynamin- and clathrin-dependent endocytosis.
  • Quantified Shiga toxin transport to the Golgi and endoplasmic reticulum by measuring the sulfation of a modified Shiga B fragment in both untreated and butyric acid-treated cells.

Main Results:

  • Endosome-to-Golgi transport of Shiga toxin is dependent on functional dynamin in both untreated and butyric acid-treated cells.
  • Butyric acid treatment increases Shiga toxin transport to the Golgi and sensitizes HeLa and BHK cells to the toxin.
  • Functional clathrin is required for Shiga toxin endosome-to-Golgi transport only in butyric acid-sensitized cells, indicating a differential regulatory role.

Conclusions:

  • Dynamin is essential for Shiga toxin retrograde transport from endosomes to the Golgi, irrespective of butyric acid treatment.
  • Clathrin's involvement in Shiga toxin transport is conditional, becoming necessary only when cells are sensitized by butyric acid.
  • These findings highlight distinct cellular pathways and regulatory mechanisms governing Shiga toxin trafficking and toxicity.

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