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Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways

BACKGROUND: The small molecule Eeyarestatin I (ESI) inhibits the endoplasmic reticulum (ER)-cytosol dislocation and subsequent degradation of ERAD (ER associated protein degradation) substrates. Toxins such as ricin and Shiga/Shiga-like toxins (SLTx) are endocytosed and trafficked to the ER. Their c...

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Autores principales: Aletrari, Mina-Olga, McKibbin, Craig, Williams, Helen, Pawar, Vidya, Pietroni, Paola, Lord, J. Michael, Flitsch, Sabine L., Whitehead, Roger, Swanton, Eileithyia, High, Stephen, Spooner, Robert A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3143184/
https://www.ncbi.nlm.nih.gov/pubmed/21799938
http://dx.doi.org/10.1371/journal.pone.0022713
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author Aletrari, Mina-Olga
McKibbin, Craig
Williams, Helen
Pawar, Vidya
Pietroni, Paola
Lord, J. Michael
Flitsch, Sabine L.
Whitehead, Roger
Swanton, Eileithyia
High, Stephen
Spooner, Robert A.
author_facet Aletrari, Mina-Olga
McKibbin, Craig
Williams, Helen
Pawar, Vidya
Pietroni, Paola
Lord, J. Michael
Flitsch, Sabine L.
Whitehead, Roger
Swanton, Eileithyia
High, Stephen
Spooner, Robert A.
author_sort Aletrari, Mina-Olga
collection PubMed
description BACKGROUND: The small molecule Eeyarestatin I (ESI) inhibits the endoplasmic reticulum (ER)-cytosol dislocation and subsequent degradation of ERAD (ER associated protein degradation) substrates. Toxins such as ricin and Shiga/Shiga-like toxins (SLTx) are endocytosed and trafficked to the ER. Their catalytic subunits are thought to utilise ERAD-like mechanisms to dislocate from the ER into the cytosol, where a proportion uncouples from the ERAD process, recovers a catalytic conformation and destroys their cellular targets. We therefore investigated ESI as a potential inhibitor of toxin dislocation. METHODOLOGY AND PRINCIPAL FINDINGS: Using cytotoxicity measurements, we found no role for ES(I) as an inhibitor of toxin dislocation from the ER, but instead found that for SLTx, ESI treatment of cells was protective by reducing the rate of toxin delivery to the ER. Microscopy of the trafficking of labelled SLTx and its B chain (lacking the toxic A chain) showed a delay in its accumulation at a peri-nuclear location, confirmed to be the Golgi by examination of SLTx B chain metabolically labelled in the trans-Golgi cisternae. The drug also reduced the rate of endosomal trafficking of diphtheria toxin, which enters the cytosol from acidified endosomes, and delayed the Golgi-specific glycan modifications and eventual plasma membrane appearance of tsO45 VSV-G protein, a classical marker for anterograde trafficking. CONCLUSIONS AND SIGNIFICANCE: ESI acts on one or more components that function during vesicular transport, whilst at least one retrograde trafficking pathway, that of ricin, remains unperturbed.
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spelling pubmed-31431842011-07-28 Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways Aletrari, Mina-Olga McKibbin, Craig Williams, Helen Pawar, Vidya Pietroni, Paola Lord, J. Michael Flitsch, Sabine L. Whitehead, Roger Swanton, Eileithyia High, Stephen Spooner, Robert A. PLoS One Research Article BACKGROUND: The small molecule Eeyarestatin I (ESI) inhibits the endoplasmic reticulum (ER)-cytosol dislocation and subsequent degradation of ERAD (ER associated protein degradation) substrates. Toxins such as ricin and Shiga/Shiga-like toxins (SLTx) are endocytosed and trafficked to the ER. Their catalytic subunits are thought to utilise ERAD-like mechanisms to dislocate from the ER into the cytosol, where a proportion uncouples from the ERAD process, recovers a catalytic conformation and destroys their cellular targets. We therefore investigated ESI as a potential inhibitor of toxin dislocation. METHODOLOGY AND PRINCIPAL FINDINGS: Using cytotoxicity measurements, we found no role for ES(I) as an inhibitor of toxin dislocation from the ER, but instead found that for SLTx, ESI treatment of cells was protective by reducing the rate of toxin delivery to the ER. Microscopy of the trafficking of labelled SLTx and its B chain (lacking the toxic A chain) showed a delay in its accumulation at a peri-nuclear location, confirmed to be the Golgi by examination of SLTx B chain metabolically labelled in the trans-Golgi cisternae. The drug also reduced the rate of endosomal trafficking of diphtheria toxin, which enters the cytosol from acidified endosomes, and delayed the Golgi-specific glycan modifications and eventual plasma membrane appearance of tsO45 VSV-G protein, a classical marker for anterograde trafficking. CONCLUSIONS AND SIGNIFICANCE: ESI acts on one or more components that function during vesicular transport, whilst at least one retrograde trafficking pathway, that of ricin, remains unperturbed. Public Library of Science 2011-07-25 /pmc/articles/PMC3143184/ /pubmed/21799938 http://dx.doi.org/10.1371/journal.pone.0022713 Text en Aletrari et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Aletrari, Mina-Olga
McKibbin, Craig
Williams, Helen
Pawar, Vidya
Pietroni, Paola
Lord, J. Michael
Flitsch, Sabine L.
Whitehead, Roger
Swanton, Eileithyia
High, Stephen
Spooner, Robert A.
Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title_full Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title_fullStr Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title_full_unstemmed Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title_short Eeyarestatin 1 Interferes with Both Retrograde and Anterograde Intracellular Trafficking Pathways
title_sort eeyarestatin 1 interferes with both retrograde and anterograde intracellular trafficking pathways
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3143184/
https://www.ncbi.nlm.nih.gov/pubmed/21799938
http://dx.doi.org/10.1371/journal.pone.0022713
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