title: The effect of acetaminophen on ubiquitin homeostasis in Saccharomyces cerevisiae.
abstract: Acetaminophen (APAP), although considered a safe drug, is one of the major causes of acute liver failure by overdose, and therapeutic chronic use can cause serious health problems. Although the reactive APAP metabolite N-acetyl-p-benzoquinoneimine (NAPQI) is clearly linked to liver toxicity, toxicity of APAP is also found without drug metabolism of APAP to NAPQI. To get more insight into mechanisms of APAP toxicity, a genome-wide screen in Saccharomyces cerevisiae for APAP-resistant deletion strains was performed. In this screen we identified genes related to the DNA damage response. Next, we investigated the link between genotype and APAP-induced toxicity or resistance by performing a more detailed screen with a library containing mutants of 1522 genes related to nuclear processes, like DNA repair and chromatin remodelling. We identified 233 strains that had an altered growth rate relative to wild type, of which 107 showed increased resistance to APAP and 126 showed increased sensitivity. Gene Ontology analysis identified ubiquitin homeostasis, regulation of transcription of RNA polymerase II genes, and the mitochondria-to-nucleus signalling pathway to be associated with APAP resistance, while histone exchange and modification, and vesicular transport were connected to APAP sensitivity. Indeed, we observed a link between ubiquitin levels and APAP resistance, whereby ubiquitin deficiency conferred resistance to APAP toxicity while ubiquitin overexpression resulted in sensitivity. The toxicity profile of various chemicals, APAP, and its positional isomer AMAP on a series of deletion strains with ubiquitin deficiency showed a unique resistance pattern for APAP. Furthermore, exposure to APAP increased the level of free ubiquitin and influenced the ubiquitination of proteins. Together, these results uncover a role for ubiquitin homeostasis in APAP-induced toxicity.
authors.affiliation: AIMMS-Division of Molecular Toxicology, Department of Chemistry and Pharmaceutical Sciences, VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Angelina
authors.initials: A
authors.lastname: Huseinovic
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Jolanda S
authors.initials: JS
authors.lastname: van Leeuwen
authors.affiliation: Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Tibor
authors.initials: T
authors.lastname: van Welsem
authors.affiliation: Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Iris
authors.initials: I
authors.lastname: Stulemeijer
authors.affiliation: Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Fred
authors.initials: F
authors.lastname: van Leeuwen
authors.affiliation: AIMMS-Division of Molecular Toxicology, Department of Chemistry and Pharmaceutical Sciences, VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Nico P E
authors.initials: NP
authors.lastname: Vermeulen
authors.affiliation: AIMMS-Department of Molecular Cell Biology, Section Genetics, VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Jan M
authors.initials: JM
authors.lastname: Kooter
authors.affiliation: AIMMS-Division of Molecular Toxicology, Department of Chemistry and Pharmaceutical Sciences, VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: J Chris
authors.initials: JC
authors.lastname: Vos
mesh_terms: D000082:Acetaminophen / Q000494:pharmacology*; D004260:DNA Repair; D017353:Gene Deletion; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D025801:Ubiquitin; D000082:Acetaminophen