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The cytotoxic effect of diphtheria toxin on the actin cytoskeleton

Cellular & Molecular Biology Letters volume 17, pages 49–61 (2012)Cite this article

Abstract

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.

Abbreviations

DT:

diphtheria toxin

FA:

fragment A

F-actin:

filamentous actin

G-actin:

globular actin monomer

HUVEC:

human umbilical vein endothelial cells

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Authors and Affiliations

  1. İstanbul Faculty of Medicine, Department of Biophysics, İstanbul University, 34390, Çapa-İstanbul, Türkiye

    Başak Varol, Muhammet Bektaş, Rüstem Nurten & Engin Bermek

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  1. Başak Varol
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  2. Muhammet Bektaş
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  4. Engin Bermek
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Correspondence to Muhammet Bektaş.

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Varol, B., Bektaş, M., Nurten, R. et al. The cytotoxic effect of diphtheria toxin on the actin cytoskeleton. Cell Mol Biol Lett 17, 49–61 (2012). https://doi.org/10.2478/s11658-011-0036-6

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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