Identification of a DNA-damage-inducible regulon in Acinetobacter baumannii
Jesús Aranda1, Margarita Poza, Miguel Shingu-Vázquez
1Servizo de Microbioloxía-INIBIC, Complexo Hospitalario Universitario A Coruña, A Coruña, Spain.
Abstract:
The transcriptional response of Acinetobacter baumannii, a major cause of nosocomial infections, to the DNA-damaging agent mitomycin C (MMC) was studied using DNA microarray technology. Most of the 39 genes induced by MMC were related to either prophages or encoded proteins involved in DNA repair. Electrophoretic mobility shift assays demonstrated that the product of the A. baumannii MMC-inducible umuD gene (umuDAb) specifically binds to the palindromic sequence TTGAAAATGTAACTTTTTCAA present in its promoter region. Mutations in this palindromic region abolished UmuDAb protein binding. A comparison of the promoter regions of all MMC-induced genes identified four additional transcriptional units with similar palindromic sequences recognized and specifically bound by UmuDAb. Therefore, the UmuDAb regulon consists of at least eight genes encoding seven predicted error-prone DNA polymerase V components and DddR, a protein of unknown function. Expression of these genes was not induced in the MMC-treated recA mutant. Furthermore, inactivation of the umuDAb gene resulted in the deregulation of all DNA-damage-induced genes containing the described palindromic DNA motif. Together, these findings suggest that UmuDAb is a direct regulator of the DNA damage response in A. baumannii.
Insights
Acinetobacter baumannii
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Acinetobacter baumannii is a significant cause of hospital-acquired infections.
- Understanding its response to DNA damage is crucial for developing new treatments.
Purpose of the Study:
- To investigate the transcriptional response of Acinetobacter baumannii to mitomycin C (MMC).
- To identify the regulatory mechanisms governing DNA damage repair pathways in this pathogen.
Main Methods:
- DNA microarray analysis to profile gene expression changes.
- Electrophoretic mobility shift assays (EMSA) to study protein-DNA interactions.
- Gene inactivation and mutation analysis.
Main Results:
- MMC induced 39 genes in A. baumannii, primarily related to prophages and DNA repair.
- The UmuDAb protein specifically binds to palindromic sequences in the promoter regions of MMC-induced genes.
- UmuDAb regulates at least eight genes, including those encoding DNA polymerase V components and DddR.
- RecA and UmuDAb are essential for the coordinated DNA damage response.
Conclusions:
- UmuDAb acts as a direct transcriptional regulator of the DNA damage response in Acinetobacter baumannii.
- The identified UmuDAb regulon provides insights into the pathogen's survival mechanisms under genotoxic stress.
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