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Eeyarestatin 1 interferes with both retrograde and anterograde intracellular trafficking pathways

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Aletrari, Mina-Olga , McKibbin, Craig, Williams, Helen, Pawar, Vidya, Pietroni, Paola, Lord, Mike (J. Mike), Flitsch, Sabine L., Whitehead, Roger C., Swanton, Eileithyia, High, Stephen and Spooner, Robert A. . (2011) Eeyarestatin 1 interferes with both retrograde and anterograde intracellular trafficking pathways. PLoS One, Vol.6 (No.7). e22713. ISSN 1932-6203

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Official URL: http://dx.doi.org/10.1371/journal.pone.0022713

Abstract

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 ESI 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.

Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology
Divisions:	Faculty of Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH):	Ricin, Endoplasmic reticulum, Biological transport
Journal or Publication Title:	PLoS One
Publisher:	Public Library of Science
ISSN:	1932-6203
Date:	25 July 2011
Volume:	Vol.6
Number:	No.7
Page Range:	e22713
Identification Number:	10.1371/journal.pone.0022713
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), National Institutes of Health (U.S.) (NIH), Wellcome Trust (London, England)
Grant number:	BB/D005752/1 (BBSRC), 5U01AI65869-02 (NIH), 080566/Z/06/Z (WT)
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URI:	http://wrap.warwick.ac.uk/id/eprint/38354