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PMID:22275523

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Citation

Wei, Q, Minh, PN, Dötsch, A, Hildebrand, F, Panmanee, W, Elfarash, A, Schulz, S, Plaisance, S, Charlier, D, Hassett, D, Häussler, S and Cornelis, P (2012) Global regulation of gene expression by OxyR in an important human opportunistic pathogen. Nucleic Acids Res. 40:4320-33

Abstract

Most bacteria control oxidative stress through the H(2)O(2)-responsive transactivator OxyR, a member of the LTTR family (LysR Type Transcriptional Regulators), which activates the expression of defensive genes such as those encoding catalases, alkyl hydroperoxide reductases and superoxide dismutases. In the human opportunistic pathogen Pseudomonas aeruginosa, OxyR positively regulates expression of the oxidative stress response genes katA, katB, ahpB and ahpCF. To identify additional targets of OxyR in P. aeruginosa PAO1, we performed chromatin immunoprecipitation in combination with whole genome tiling array analyses (ChIP-chip). We detected 56 genes including all the previously identified defensive genes and a battery of novel direct targets of OxyR. Electrophoretic mobility shift assays (EMSAs) for selected newly identified targets indicated that ∼70% of those were bound by purified oxidized OxyR and their regulation was confirmed by quantitative real-time polymerase chain reaction. Furthermore, a thioredoxin system was identified to enzymatically reduce OxyR under oxidative stress. Functional classification analysis showed that OxyR controls a core regulon of oxidative stress defensive genes, and other genes involved in regulation of iron homeostasis (pvdS), quorum-sensing (rsaL), protein synthesis (rpsL) and oxidative phosphorylation (cyoA and snr1). Collectively, our results indicate that OxyR is involved in oxidative stress defense and regulates other aspects of cellular metabolism as well.

Links

PubMed PMC3378865 Online version:10.1093/nar/gks017

Keywords

Binding Sites; Chromatin Immunoprecipitation; Gene Expression Regulation, Bacterial; Genome, Bacterial; Oligonucleotide Array Sequence Analysis; Oxidative Stress/genetics; Pseudomonas aeruginosa/enzymology; Pseudomonas aeruginosa/genetics; Pseudomonas aeruginosa/metabolism; Regulon; Thioredoxins/metabolism; Trans-Activators/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

PSEAE:Q9HTL4

enables

GO:0000976: transcription regulatory region sequence-specific DNA binding

ECO:0001807: electrophoretic mobility shift assay evidence used in manual assertion

F

Seeded From UniProt

complete

PSEAE:Q9HTL4

part_of

GO:0032993: protein-DNA complex

ECO:0001807: electrophoretic mobility shift assay evidence used in manual assertion

C

Seeded From UniProt

complete

PSEAE:Q9HTL4

enables

GO:0000976: transcription regulatory region sequence-specific DNA binding

ECO:0006007: chromatin immunoprecipitation-chip evidence used in manual assertion

F

Seeded From UniProt

complete

PSEAE:Q9HTL4

involved_in

GO:0006355: regulation of transcription, DNA-templated

ECO:0001808: reverse transcription polymerase chain reaction evidence used in manual assertion

P

Seeded From UniProt

complete

PSEAE:Q9HTL4

GO:0001071: nucleic acid binding transcription factor activity

ECO:0000314:

F

EMSA results in figure 2 prove OxyR binds to the promoter regions of several P. aerugenosa genes. Additionally, figure 3 shows that expression of dps, pvdS and trxB2 genes increased in the presence of OxyR activated by H2O2.

complete
CACAO 7589

PSEAE:Q9HTL4

part_of

GO:0032993: protein-DNA complex

ECO:0006007: chromatin immunoprecipitation-chip evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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