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 hdl:20.500.11880/29973 http://dx.doi.org/10.22028/D291-27875 |
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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toxins-08-00366.pdf | 1,52 MB | Adobe PDF | View/Open |
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