Several protein toxins, including the A chain of the plant protein ricin (RTA), enter mammalian cells by endocytosis and catalytically modify cellular components to disrupt essential cellular processes. In the case of ricin, the process inhibited is protein synthesis. In order to reach their cytosolic substrates, several toxins undergo retrograde transport to the ER (endoplasmic reticulum) before translocating across the ER membrane. To achieve this export, these toxins exploit the ERAD (ER-associated protein degradation) pathway but must escape, at least in part, the normal degradative fate of ERAD substrates in order to intoxicate the cell. Toxins that translocate from the ER have an unusually low lysine content that reduces the likelihood of ubiquitination and ubiquitin-mediated proteasomal degradation. We have changed the two lysyl residues normally present in RTA to arginyl residues. Their replacement in RTA did not have a significant stabilizing effect on the protein, suggesting that the endogenous lysyl residues are not sites for ubiquitin attachment. However, when four additional lysyl residues were introduced into RTA in a way that did not compromise the activity, structure or stability of the toxin, degradation was significantly enhanced. Enhanced degradation resulted from ubiquitination that predisposed the toxin to proteasomal degradation. Treatment with the proteasomal inhibitor lactacystin increased the cytotoxicity of the lysine-enriched RTA to a level approaching that of wild-type RTA.
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December 2003
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Conference Article|
December 01 2003
Retrograde transport of toxins across the endoplasmic reticulum membrane
J.M. Lord;
J.M. Lord
1
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
1To whom correspondence should be addressed (e-mail mlord@bio.warwick.ac.uk).
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E. Deeks;
E. Deeks
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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C.J. Marsden;
C.J. Marsden
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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K. Moore;
K. Moore
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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C. Pateman;
C. Pateman
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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D.C. Smith;
D.C. Smith
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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R.A. Spooner;
R.A. Spooner
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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P. Watson;
P. Watson
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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L.M. Roberts
L.M. Roberts
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2003 Biochemical Society
2003
Biochem Soc Trans (2003) 31 (6): 1260–1262.
Citation
J.M. Lord, E. Deeks, C.J. Marsden, K. Moore, C. Pateman, D.C. Smith, R.A. Spooner, P. Watson, L.M. Roberts; Retrograde transport of toxins across the endoplasmic reticulum membrane. Biochem Soc Trans 1 December 2003; 31 (6): 1260–1262. doi: https://doi.org/10.1042/bst0311260
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