title: Phenotype enhancement screen of a regulatory spx mutant unveils a role for the ytpQ gene in the control of iron homeostasis.
abstract: Spx is a global regulator of genes that are induced by disulfide stress in Bacillus subtilis. The regulon that it governs is comprised of over 120 genes based on microarray analysis, although it is not known how many of these are under direct Spx control. Most of the Spx-regulated genes (SRGs) are of unknown function, but many encode products that are conserved in low %GC Gram-positive bacteria. Using a gene-disruption library of B. subtilis genomic mutations, the SRGs were screened for phenotypes related to Spx-controlled activities, such as poor growth in minimal medium and sensitivity to methyglyoxal, but nearly all of the SRG mutations showed little if any phenotype. To uncover SRG function, the mutations were rescreened in an spx mutant background to determine which mutant SRG allele would enhance the spx mutant phenotype. One of the SRGs, ytpQ was the site of a mutation that, when combined with an spx null mutation, elevated the severity of the Spx mutant phenotype, as shown by reduced growth in a minimal medium and by hypersensitivity to methyglyoxal. The ytpQ mutant showed elevated oxidative protein damage when exposed to methylglyoxal, and reduced growth rate in liquid culture. Proteomic and transcriptomic data indicated that the ytpQ mutation caused the derepression of the Fur and PerR regulons of B. subtilis. Our study suggests that the ytpQ gene, encoding a conserved DUF1444 protein, functions directly or indirectly in iron homeostasis. The ytpQ mutant phenotype mimics that of a fur mutation, suggesting a condition of low cellular iron. In vitro transcription analysis indicated that Spx stimulates transcription from the ytpPQR operon within which the ytpQ gene resides. The work uncovers a link between Spx and control of iron homeostasis.
authors.affiliation: Division of Environmental and Biomolecular Systems, Institute of Environmental Health, Oregon Health and Science University, Beaverton, Oregon, United States of America. pzuber@ebs.ogi.edu
authors.forename: Peter
authors.initials: P
authors.lastname: Zuber
authors.forename: Shefali
authors.initials: S
authors.lastname: Chauhan
authors.forename: Praseeda
authors.initials: P
authors.lastname: Pilaka
authors.forename: Michiko M
authors.initials: MM
authors.lastname: Nakano
authors.forename: Sairam
authors.initials: S
authors.lastname: Gurumoorthy
authors.forename: Ann A
authors.initials: AA
authors.lastname: Lin
authors.forename: Skye M
authors.initials: SM
authors.lastname: Barendt
authors.forename: Bui Khanh
authors.initials: BK
authors.lastname: Chi
authors.forename: Haike
authors.initials: H
authors.lastname: Antelmann
authors.forename: Ulrike
authors.initials: U
authors.lastname: Mäder
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D020869:Gene Expression Profiling; D005809:Genes, Regulator / Q000235:genetics*; D005816:Genetic Complementation Test; D006706:Homeostasis; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D009876:Operon / Q000235:genetics*; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000235:genetics; D040901:Proteomics; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012097:Repressor Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D015415:Biomarkers; D004220:Disulfides; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; D007501:Iron