Cargando…
The cytotoxic effect of diphtheria toxin on the actin cytoskeleton
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 depolymeri...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SP Versita
2011
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275567/ https://www.ncbi.nlm.nih.gov/pubmed/22139586 http://dx.doi.org/10.2478/s11658-011-0036-6 |
_version_ | 1783377828531666944 |
---|---|
author | Varol, Başak Bektaş, Muhammet Nurten, Rüstem Bermek, Engin |
author_facet | Varol, Başak Bektaş, Muhammet Nurten, Rüstem Bermek, Engin |
author_sort | Varol, Başak |
collection | PubMed |
description | 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 K(m) = 2.2 nM; V(max) = 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. |
format | Online Article Text |
id | pubmed-6275567 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | SP Versita |
record_format | MEDLINE/PubMed |
spelling | pubmed-62755672018-12-10 The cytotoxic effect of diphtheria toxin on the actin cytoskeleton Varol, Başak Bektaş, Muhammet Nurten, Rüstem Bermek, Engin Cell Mol Biol Lett Research Article 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 K(m) = 2.2 nM; V(max) = 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. SP Versita 2011-12-02 /pmc/articles/PMC6275567/ /pubmed/22139586 http://dx.doi.org/10.2478/s11658-011-0036-6 Text en © © Versita Warsaw and Springer-Verlag Wien 2011 |
spellingShingle | Research Article Varol, Başak Bektaş, Muhammet Nurten, Rüstem Bermek, Engin The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title | The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title_full | The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title_fullStr | The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title_full_unstemmed | The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title_short | The cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
title_sort | cytotoxic effect of diphtheria toxin on the actin cytoskeleton |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275567/ https://www.ncbi.nlm.nih.gov/pubmed/22139586 http://dx.doi.org/10.2478/s11658-011-0036-6 |
work_keys_str_mv | AT varolbasak thecytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT bektasmuhammet thecytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT nurtenrustem thecytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT bermekengin thecytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT varolbasak cytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT bektasmuhammet cytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT nurtenrustem cytotoxiceffectofdiphtheriatoxinontheactincytoskeleton AT bermekengin cytotoxiceffectofdiphtheriatoxinontheactincytoskeleton |