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Communication

Yeast Reporter Assay to Identify Cellular Components of Ricin Toxin A Chain Trafficking

Molecular and Cell Biology, Department of Biosciences and Center of Human and Molecular Biology (ZHMB), Saarland University, Saarbrücken D-66123, Germany
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Academic Editor: Tomas Girbes
Toxins 2016, 8(12), 366; https://doi.org/10.3390/toxins8120366
Received: 11 October 2016 / Revised: 22 November 2016 / Accepted: 30 November 2016 / Published: 6 December 2016
(This article belongs to the Special Issue Ribosome Inactivating Toxins)
RTA, the catalytic A-subunit of the ribosome inactivating A/B toxin ricin, inhibits eukaryotic protein biosynthesis by depurination of 28S rRNA. Although cell surface binding of ricin holotoxin is mainly mediated through its B-subunit (RTB), sole application of RTA is also toxic, albeit to a significantly lower extent, suggesting alternative pathways for toxin uptake and transport. Since ricin toxin trafficking in mammalian cells is still not fully understood, we developed a GFP-based reporter assay in yeast that allows rapid identification of cellular components required for RTA uptake and subsequent transport through a target cell. We hereby show that Ypt6p, Sft2p and GARP-complex components play an important role in RTA transport, while neither the retromer complex nor COPIB vesicles are part of the transport machinery. Analyses of yeast knock-out mutants with chromosomal deletion in genes whose products regulate ADP-ribosylation factor GTPases (Arf-GTPases) and/or retrograde Golgi-to-ER (endoplasmic reticulum) transport identified Sso1p, Snc1p, Rer1p, Sec22p, Erv46p, Gea1p and Glo3p as novel components in RTA transport, suggesting the developed reporter assay as a powerful tool to dissect the multistep processes of host cell intoxication in yeast. View Full-Text
Keywords: S. cerevisiae; ricin toxin A chain (RTA); ribosome inactivating protein (RIP); retrograde protein transport; trans-Golgi network (TGN); endoplasmic reticulum (ER) S. cerevisiae; ricin toxin A chain (RTA); ribosome inactivating protein (RIP); retrograde protein transport; trans-Golgi network (TGN); endoplasmic reticulum (ER)
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MDPI and ACS Style

Becker, B.; Schnöder, T.; Schmitt, M.J. Yeast Reporter Assay to Identify Cellular Components of Ricin Toxin A Chain Trafficking. Toxins 2016, 8, 366. https://doi.org/10.3390/toxins8120366

AMA Style

Becker B, Schnöder T, Schmitt MJ. Yeast Reporter Assay to Identify Cellular Components of Ricin Toxin A Chain Trafficking. Toxins. 2016; 8(12):366. https://doi.org/10.3390/toxins8120366

Chicago/Turabian Style

Becker, Björn, Tina Schnöder, and Manfred J. Schmitt. 2016. "Yeast Reporter Assay to Identify Cellular Components of Ricin Toxin A Chain Trafficking" Toxins 8, no. 12: 366. https://doi.org/10.3390/toxins8120366

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