Open Access

The cytotoxic effect of diphtheria toxin on the actin cytoskeleton

  • Başak Varol1,
  • Muhammet Bektaş1Email author,
  • Rüstem Nurten1 and
  • Engin Bermek1
Cellular & Molecular Biology LettersAn International Journal201117:36

DOI: 10.2478/s11658-011-0036-6

Received: 11 June 2011

Accepted: 23 November 2011

Published: 2 December 2011


Diphtheria toxin (DT) and its N-terminal fragment A (FA) catalyse the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide (NAD) into a covalent linkage with eukaryotic elongation factor 2 (eEF2). DT-induced cytotoxicity is versatile, and it includes DNA cleavage and the depolymerisation of actin filaments. The inhibition of the ADP-ribosyltransferase (ADPrT) activity of FA did not affect the deoxyribonuclease activity of FA or its interaction with actin. The toxin entry rate into cells (HUVEC) was determined by measuring the ADP-ribosyltransferase activity. DT uptake was nearly 80% after 30 min. The efficiency was determined as Km = 2.2 nM; Vmax = 0.25 pmol.min−1. The nuclease activity was tested with hyperchromicity experiments, and it was concluded that G-actin has an inhibitory effect on DT nuclease activity. In thepresence of DT and mutant of diphtheria toxin (CRM197), F-actin depolymerisation was determined with gel filtration, WB and fluorescence techniques. In the presence of DT and CRM197, 60–65% F-actin depolymerisation was observed. An in vitro FA-actin interaction and F-actin depolymerisation were reported in our previous paper. The present study thus confirms the depolymerisation of actin cytoskeleton in vivo.

Key words

ADP-ribosylation Diphtheria toxin Eukaryotic elongation factor-2 F-actin Fragment A