title: Identification of an overexpressed yeast gene which prevents aminoglycoside toxicity.
abstract: Aminoglycoside antibiotics, used to treat bacterial infections by interfering with proofreading during protein synthesis, cause sensorineural hearing loss in genetically susceptible individuals. The only aminoglycoside-hypersensitivity mutations which have been described in humans are in the mitochondrial 125 rRNA gene, potentially allowing increased antibiotic binding to mitochondrial ribosomes. To identify additional predisposing mutations, a yeast model system was used to isolate genes which interact with or bypass the effects of aminoglycoside antibiotics. A novel yeast gene was isolated which, in high copy, confers neomycin resistance to yeast transformants. The neomycin-resistance 1 gene (NEO1) encodes a potential 1151 as integral membrane protein, most homologous to the yeast DRS2 gene product, a Ca(2+)-ATPase involved in cytoplasmic ribosome assembly. The N-terminus of Neo1p is partially homologous to abrin A-chain, another protein which interacts with cytoplasmic ribosomes. Mutagenesis experiments demonstrate that the NEO1 product is essential for vegetative growth and that the drug-resistance phenotype requires ATPase function.
authors.affiliation: Ahmanson Department of Pediatrics, Steven Spielberg Pediatric Research Center, Los Angeles, CA, USA. tPrezant@mailgate.csmc.edu
authors.forename: T R
authors.initials: TR
authors.lastname: Prezant
authors.forename: W E
authors.initials: WE
authors.lastname: Chaltraw
authors.forename: N
authors.initials: N
authors.lastname: Fischel-Ghodsian
mesh_terms: D000251:Adenosine Triphosphatases; D000595:Amino Acid Sequence; D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000633:toxicity*; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000096:biosynthesis / Q000737:chemistry; D017124:Conserved Sequence; D017931:DNA Primers; D004352:Drug Resistance, Microbial / Q000235:genetics*; D005800:Genes, Fungal; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D049294:Phospholipid Transfer Proteins; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D017931:DNA Primers; C104014:DRS2 protein, S cerevisiae; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D049294:Phospholipid Transfer Proteins; D029701:Saccharomyces cerevisiae Proteins; D000251:Adenosine Triphosphatases; D000252:Calcium-Transporting ATPases; C104013:NEO1 protein, S cerevisiae