A novel, drug-based, cellular assay for the activity of neurotoxic mutants of the prion protein.
In prion diseases, the infectious isoform of the prion protein (PrP(Sc)) may subvert a normal, physiological activity of the cellular isoform (PrP(C)). A deletion mutant of the prion protein (Delta105-125) that produces a neonatal lethal phenotype when expressed in transgenic mice provides a window into the normal function of PrP(C) and how it can be corrupted to produce neurotoxic effects. We report here the surprising and unexpected observation that cells expressing Delta105-125 PrP and related mutants are hypersensitive to the toxic effects of two classes of antibiotics (aminoglycosides and bleomycin analogues) that are commonly used for selection of stably transfected cell lines. This unusual phenomenon mimics several essential features of Delta105-125 PrP toxicity seen in transgenic mice, including rescue by co-expression of wild type PrP. Cells expressing Delta105-125 PrP are susceptible to drug toxicity within minutes, suggesting that the mutant protein enhances cellular accumulation of these cationic compounds. Our results establish a screenable cellular phenotype for the activity of neurotoxic forms of PrP, and they suggest possible mechanisms by which these molecules could produce their pathological effects in vivo.
Massignan
Tania
T
Department of Cell Biology and Physiology, Washington University School of Medicine, St Louis, Missouri 63110, USA.
Stewart
Richard S
RS
Biasini
Emiliano
E
Solomon
Isaac H
IH
Bonetto
Valentina
V
Chiesa
Roberto
R
Harris
David A
DA
D000563:Amebicides / Q000494:pharmacology; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods*; D001761:Bleomycin / Q000494:pharmacology; D002460:Cell Line / Q000187:drug effects; D002470:Cell Survival; D002934:Cinnamates / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D058851:GPI-Linked Proteins; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D010641:Phenotype; D018621:PrPC Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D018620:PrPSc Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D017096:Prion Diseases / Q000378:metabolism*; D011328:Prions / Q000235:genetics / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology
D000563:Amebicides; D000903:Antibiotics, Antineoplastic; D002934:Cinnamates; D058851:GPI-Linked Proteins; D005839:Gentamicins; C501125:PRND protein, human; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D020033:Protein Isoforms; D001761:Bleomycin; C105427:Zeocin; D006921:Hygromycin B; C026273:hygromycin A; C010680:antibiotic G 418