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Titel: Adapting Yeast as Model to Study Ricin Toxin A Uptake and Trafficking
VerfasserIn: Becker, Björn
Schmitt, Manfred J.
Sprache: Englisch
Titel: Toxins
Bandnummer: 3
Heft: 7
Startseite: 834
Endseite: 847
Verlag/Plattform: MDPI
Erscheinungsjahr: 2011
Freie Schlagwörter: ricin toxin A chain
ER retention signal
yeast spheroplasts
toxin endocytosis and transport
oyxgen-sensor microtiter plate
H/KDEL
DDC-Sachgruppe: 570 Biowissenschaften, Biologie
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: The plant A/B toxin ricin represents a heterodimeric glycoprotein belonging to the family of ribosome inactivating proteins, RIPs. Its toxicity towards eukaryotic cells results from the depurination of 28S rRNA due to the N-glycosidic activity of ricin toxin A chain, RTA. Since the extention of RTA by a mammalian-specific endoplasmic reticulum (ER) retention signal (KDEL) significantly increases RTA in vivo toxicity against mammalian cells, we here analyzed the phenotypic effect of RTA carrying the yeast-specific ER retention motif HDEL. Interestingly, such a toxin (RTAHDEL) showed a similar cytotoxic effect on yeast as a corresponding RTAKDEL variant on HeLa cells. Furthermore, we established a powerful yeast bioassay for RTA in vivo uptake and trafficking which is based on the measurement of dissolved oxygen in toxin-treated spheroplast cultures of S. cerevisiae. We show that yeast spheroplasts are highly sensitive against external applied RTA and further demonstrate that its toxicity is greatly enhanced by replacing the C-terminal KDEL motif by HDEL. Based on the RTA resistant phenotype seen in yeast knock-out mutants defective in early steps of endocytosis (∆end3) and/or in RTA depurination activity on 28S rRNA (∆rpl12B) we feel that the yeast-based bioassay described in this study is a powerful tool to dissect intracellular A/B toxin transport from the plasma membrane through the endosomal compartment to the ER.
DOI der Erstveröffentlichung: 10.3390/toxins3070834
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-278694
hdl:20.500.11880/29969
http://dx.doi.org/10.22028/D291-27869
ISSN: 2072-6651
Datum des Eintrags: 6-Nov-2020
Fakultät: NT - Naturwissenschaftlich- Technische Fakultät
Fachrichtung: NT - Biowissenschaften
Professur: NT - Prof. Dr. Manfred Schmitt
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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