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| Status |
Public on Oct 25, 2011 |
| Title |
Phenotype Enhancement Screen of a Regulatory spx Mutant Unveils a Role for the ytpQ Gene in the Control of Iron Homeostasis. |
| Platform organism |
Bacillus subtilis subsp. subtilis str. 168 |
| Sample organism |
Bacillus subtilis |
| Experiment type |
Expression profiling by array
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| Summary |
Spx is a global regulator of genes that are induced by disulfide stress in Bacillus subtilis. Most of the Spx-regulated genes (SRGs) are of unknown function, but many encode products 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 growth in minimal medium and sensitivity to methylglyoxal, 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 methylglyoxal. Proteomic and transcriptomic data indicated that the ytpQ mutation caused the derepression of the Fur and PerR regulons, as well as heightened LexA-controlled gene expression. Our study suggests that the ytpQ gene, encoding a conserved DUF1444 protein, functions directly or indirectly in repairing or stabilizing Fe-bearing proteins, which are sensitive to thiol reactive agents and, therefore, likely influenced by Spx-controlled genes.
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| Overall design |
The wild type parent, ytpQ and spx mutants were grown in a glucose minimal medium, with and without 2.8 mM methylglyoxal, to mid-log phase. Cells were harvested and RNA was extracted for microarray hybridization analysis to determine if any changes in the transcriptome could be detected that were attributable to the ytpQ mutation. Microarray hybridizations were performed with RNA from three biological replicates.
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| Contributor(s) |
Chauhan S, Pilaka P, Nakano MM, Gurumoorthy S, Chi BK, Mäder U, Antelmann H, Zuber P |
| Citation(s) |
21949854 |
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| Submission date |
Apr 26, 2011 |
| Last update date |
Apr 06, 2016 |
| Contact name |
Ulrike Mäder |
| E-mail(s) |
[email protected]
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| Organization name |
University Medicine Greifswald
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| Department |
Functional Genomics
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| Street address |
F.-L.-Jahn-Str. 15A
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| City |
Greifswald |
| ZIP/Postal code |
D-17489 |
| Country |
Germany |
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| Platforms (1) |
| GPL10901 |
Bacillus subtilis Custom Agilent 44k v2 |
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| Samples (9)
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| GSM714792 |
ytpQ mutant, methylglyoxal (MG) treated, rep1 |
| GSM714793 |
ytpQ mutant, methylglyoxal (MG) treated, rep2 |
| GSM714794 |
ytpQ mutant, methylglyoxal (MG) treated, rep3 |
| GSM714795 |
spx mutant, untreated, rep1 |
| GSM714796 |
spx mutant, untreated, rep2 |
| GSM714797 |
spx mutant, untreated, rep3 |
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| Relations |
| BioProject |
PRJNA140593 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE28872_RAW.tar |
121.1 Mb |
(http)(custom) |
TAR (of TXT) |
| GSE28872_ytpQ_and_spx_annotated_genes.txt.gz |
144.7 Kb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data are available on Series record |
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