RP - Screen to identify new components in ERQD (»Add to Infobox)

Wolf Dieter H.

Dieter H. Wolf's Group

For identification of new components involved in endoplasmic reticulum quality control and associated protein degradation (ERQD), an available yeast mutant library is used for a genome-wide screen, which involves approximately 5000 mutant strains each deleted in a single nonessential gene (EUROSCARF, Frankfurt, Germany). Use of this genomic library is possible, because cells defective in ERQD can tolerate defects in this process as long as the unfolded protein response (UPR) is intact (Friedländer et al., 2000; Travers et al., 2000). A plasmid encoding the well characterized ERQD substrate CPY* fused to a transmembrane domain and the marker protein LEU2, called CTL*, is transformed into all 5000 mutants. In case of wild-type degradation of this chimeric protein, the cells are not able to grow on medium lacking the complementing nutrient. In contrast, disturbance of the ERQD pathway prevents the degradation of the ERQD substrate carrying the marker protein fusion and allows cell growth under selective conditions (Schäfer and Wolf, 2005).

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Wolf Dieter H. (Principal Investigator / Dieter H. Wolf's Group)

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