title: Saccharomyces cerevisiae genome-wide mutant screen for sensitivity to 2,4-diacetylphloroglucinol, an antibiotic produced by Pseudomonas fluorescens.
abstract: 2,4-Diacetylphloroglucinol (2,4-DAPG), an antibiotic produced by Pseudomonas fluorescens, has broad-spectrum antibiotic activity, inhibiting organisms ranging from viruses, bacteria, and fungi to higher plants and mammalian cells. The biosynthesis and regulation of 2,4-DAPG in P. fluorescens are well described, but the mode of action against target organisms is poorly understood. As a first step to elucidate the mechanism, we screened a deletion library of Saccharomyces cerevisiae in broth and agar medium supplemented with 2,4-DAPG. We identified 231 mutants that showed increased sensitivity to 2,4-DAPG under both conditions, including 22 multidrug resistance-related mutants. Three major physiological functions correlated with an increase in sensitivity to 2,4-DAPG: membrane function, reactive oxygen regulation, and cell homeostasis. Physiological studies with wild-type yeast validated the results of the mutant screens. The chemical-genetic fitness profile of 2,4-DAPG resembled those of menthol, sodium azide, and hydrogen peroxide determined in previous high-throughput screening studies. Collectively, these findings indicate that 2,4-DAPG acts on multiple basic cellular processes.
authors.affiliation: P.O. Box 646430, 367 Johnson Hall, Washington State University, Pullman, WA 99164-6430, USA.
authors.forename: Youn-Sig
authors.initials: YS
authors.lastname: Kwak
authors.forename: Sangjo
authors.initials: S
authors.lastname: Han
authors.forename: Linda S
authors.initials: LS
authors.lastname: Thomashow
authors.forename: Jennifer T
authors.initials: JT
authors.lastname: Rice
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Paulitz
authors.forename: Dongsup
authors.initials: D
authors.lastname: Kim
authors.forename: David M
authors.initials: DM
authors.lastname: Weller
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism / Q000494:pharmacology*; D017353:Gene Deletion; D010696:Phloroglucinol / Q000031:analogs & derivatives / Q000494:pharmacology; D011551:Pseudomonas fluorescens / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D000890:Anti-Infective Agents; D029701:Saccharomyces cerevisiae Proteins; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol