Response of Staphylococcus aureus to subinhibitory concentrations of a sequence-selective, DNA minor groove cross-linking pyrrolobenzodiazepine dimer
Response of Staphylococcus aureus to subinhibitory concentrations of a sequence-selective, DNA minor groove cross-linking pyrrolobenzodiazepine dimer
Objectives ELB-21 is a pyrrolo[2,1-c][1,4]benzodiazepine dimer with potent antistaphylococcal activity; it binds covalently to guanine residues on opposing strands of duplex DNA, interfering with regulatory proteins and transcription elongation in a sequence-selective manner. Transcriptional and proteomic alterations induced by exposure of Staphylococcus aureus clinical isolate EMRSA-16 to ELB-21 were determined in order to define more precisely the bactericidal mechanism of the drug.
Methods DNase I footprinting was used to identify high-affinity DNA binding sites. Microarrays and gel electrophoresis were used to assess the ELB-21-induced phenotype.
Results High-affinity interstrand binding sites in which guanine residues were separated by 4 bp, and also some intrastrand cross-linking sites of variable length were identified. Exposure of EMRSA-16 to 0.015 mg/L ELB-21 elicited a 2-fold or greater up-regulation of 168 genes in logarithmic phase and 181 genes in stationary phase; the majority of genes affected were associated with resident prophages ?Sa2 and ?Sa3, pathogenicity island SaPI4 and DNA damage repair. ELB-21 induced a marked increase in the number of viable phage particles in culture supernatants. The expression of only a limited number of genes showed a >50% reduction. Sixteen extracellular and four intracellular proteins were differentially expressed during logarithmic and stationary phases, including RecA, proteins associated with staphylococcal pathogenesis (IsaA, CspA), cell division and wall synthesis.
Conclusions ELB-21 kills S. aureus by forming multiple interstrand and intrastrand DNA cross-links, resulting in induction of the DNA damage response, derepression of resident prophages and modulation of a limited number of genes involved with cell wall synthesis.
949-959
Doyle, M.
ff930c70-b241-455a-b270-a1a67e8993fb
Feuerbaum, E.-A.
af0b614f-8643-4161-92c7-6773d4d96b25
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Hinds, J.
9e2ebf05-3e45-43c0-bda2-a0b7dd6c57b8
Thurston, D. E.
1a94ee38-b404-4cbb-9480-75697d6bbc43
Taylor, P. W.
018c97e7-10de-437d-bf5b-6a7c170c5446
10 August 2009
Doyle, M.
ff930c70-b241-455a-b270-a1a67e8993fb
Feuerbaum, E.-A.
af0b614f-8643-4161-92c7-6773d4d96b25
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Hinds, J.
9e2ebf05-3e45-43c0-bda2-a0b7dd6c57b8
Thurston, D. E.
1a94ee38-b404-4cbb-9480-75697d6bbc43
Taylor, P. W.
018c97e7-10de-437d-bf5b-6a7c170c5446
Doyle, M., Feuerbaum, E.-A., Fox, Keith R., Hinds, J., Thurston, D. E. and Taylor, P. W.
(2009)
Response of Staphylococcus aureus to subinhibitory concentrations of a sequence-selective, DNA minor groove cross-linking pyrrolobenzodiazepine dimer.
Journal of Antimicrobial Chemotherapy, 64 (5), .
(doi:10.1093/jac/dkp325).
Abstract
Objectives ELB-21 is a pyrrolo[2,1-c][1,4]benzodiazepine dimer with potent antistaphylococcal activity; it binds covalently to guanine residues on opposing strands of duplex DNA, interfering with regulatory proteins and transcription elongation in a sequence-selective manner. Transcriptional and proteomic alterations induced by exposure of Staphylococcus aureus clinical isolate EMRSA-16 to ELB-21 were determined in order to define more precisely the bactericidal mechanism of the drug.
Methods DNase I footprinting was used to identify high-affinity DNA binding sites. Microarrays and gel electrophoresis were used to assess the ELB-21-induced phenotype.
Results High-affinity interstrand binding sites in which guanine residues were separated by 4 bp, and also some intrastrand cross-linking sites of variable length were identified. Exposure of EMRSA-16 to 0.015 mg/L ELB-21 elicited a 2-fold or greater up-regulation of 168 genes in logarithmic phase and 181 genes in stationary phase; the majority of genes affected were associated with resident prophages ?Sa2 and ?Sa3, pathogenicity island SaPI4 and DNA damage repair. ELB-21 induced a marked increase in the number of viable phage particles in culture supernatants. The expression of only a limited number of genes showed a >50% reduction. Sixteen extracellular and four intracellular proteins were differentially expressed during logarithmic and stationary phases, including RecA, proteins associated with staphylococcal pathogenesis (IsaA, CspA), cell division and wall synthesis.
Conclusions ELB-21 kills S. aureus by forming multiple interstrand and intrastrand DNA cross-links, resulting in induction of the DNA damage response, derepression of resident prophages and modulation of a limited number of genes involved with cell wall synthesis.
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Published date: 10 August 2009
Organisations:
Molecular and Cellular
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Local EPrints ID: 206543
URI: http://eprints.soton.ac.uk/id/eprint/206543
ISSN: 0305-7453
PURE UUID: e9d3029b-c4b4-41a9-9a24-4c88c2d17616
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Date deposited: 22 Dec 2011 15:31
Last modified: 15 Mar 2024 02:36
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Author:
M. Doyle
Author:
E.-A. Feuerbaum
Author:
J. Hinds
Author:
D. E. Thurston
Author:
P. W. Taylor
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