Please use this identifier to cite or link to this item: doi:10.22028/D291-27875
Title: Yeast Reporter Assay to Identify Cellular Components of Ricin Toxin A Chain Trafficking
Author(s): Becker, Björn
Schnöder, Tina
Schmitt, Manfred J.
Language: English
Title: Toxins
Volume: 8
Issue: 12
Publisher/Platform: MDPI
Year of Publication: 2016
Free key words: S. cerevisiae
ricin toxin A chain (RTA)
ribosome inactivating protein (RIP)
retrograde protein transport
trans-Golgi network (TGN)
endoplasmic reticulum (ER)
DDC notations: 570 Life sciences, biology
Publikation type: Journal Article
Abstract: 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.
DOI of the first publication: 10.3390/toxins8120366
Link to this record: urn:nbn:de:bsz:291--ds-278755
ISSN: 2072-6651
Date of registration: 6-Nov-2020
Faculty: NT - Naturwissenschaftlich- Technische Fakultät
Department: NT - Biowissenschaften
Professorship: NT - Prof. Dr. Manfred Schmitt
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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