A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase
| Autor: | Jing Kang, Robert Mintzer, Kimberly Kajihara, Wouter L. W. Hazenbos, Shailesh V. Date, Peter A. S. Smith, J. Hiroshi Morisaki, Eric M. Kofoed, Man-Wah Tan, Eric J. Brown, Tommy K. Cheung, Donghong Yan, Chau Linda Truong, David Arnott, Ishita M. Shah, Michael F. T. Koehler, Zora Modrusan, Min Xu, Christopher E. Heise |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Signal peptide Staphylococcus aureus Applied Microbiology bacterial secretion 030106 microbiology Gene Expression ATP-binding cassette transporter Microbial Sensitivity Tests Biology medicine.disease_cause Microbiology Mice 03 medical and health sciences Virology Drug Resistance Bacterial Gene expression Genetics medicine Animals Secretion Selection Genetic Molecular Biology Gene Mutation Signal peptidase Virulence Serine Endopeptidases Membrane Proteins Staphylococcal Infections bacterial resistance QR1-502 In vitro Anti-Bacterial Agents Cell biology Disease Models Animal Biochemistry signal peptidase ATP-Binding Cassette Transporters Research Article |
| Zdroj: | mBio, Vol 7, Iss 5 (2016) Microbial Cell mBio, Vol 7, Iss 5, p e00412-16 (2016) mBio |
| ISSN: | 2150-7511 2161-2129 |
| DOI: | 10.1128/mbio.00412-16 |
| Popis: | The type I signal peptidase of Staphylococcus aureus, SpsB, is an attractive antibacterial target because it is essential for viability and extracellularly accessible. We synthesized compound 103, a novel arylomycin-derived inhibitor of SpsB with significant potency against various clinical S. aureus strains (MIC of ~1 µg/ml). The predominant clinical strain USA300 developed spontaneous resistance to compound 103 with high frequency, resulting from single point mutations inside or immediately upstream of cro/cI, a homolog of the lambda phage transcriptional repressor cro. These cro/cI mutations led to marked (>50-fold) overexpression of three genes encoding a putative ABC transporter. Overexpression of this ABC transporter was both necessary and sufficient for resistance and, notably, circumvented the essentiality of SpsB during in vitro culture. Mutation of its predicted ATPase gene abolished resistance, suggesting a possible role for active transport; in these bacteria, resistance to compound 103 occurred with low frequency and through mutations in spsB. Bacteria overexpressing the ABC transporter and lacking SpsB were capable of secreting a subset of proteins that are normally cleaved by SpsB and instead were cleaved at a site distinct from the canonical signal peptide. These bacteria secreted reduced levels of virulence-associated proteins and were unable to establish infection in mice. This study reveals the mechanism of resistance to a novel arylomycin derivative and demonstrates that the nominal essentiality of the S. aureus signal peptidase can be circumvented by the upregulation of a putative ABC transporter in vitro but not in vivo. IMPORTANCE The type I signal peptidase of Staphylococcus aureus (SpsB) enables the secretion of numerous proteins by cleavage of the signal peptide. We synthesized an SpsB inhibitor with potent activity against various clinical S. aureus strains. The predominant S. aureus strain USA300 develops resistance to this inhibitor by mutations in a novel transcriptional repressor (cro/cI), causing overexpression of a putative ABC transporter. This mechanism promotes the cleavage and secretion of various proteins independently of SpsB and compensates for the requirement of SpsB for viability in vitro. However, bacteria overexpressing the ABC transporter and lacking SpsB secrete reduced levels of virulence-associated proteins and are unable to infect mice. This study describes a bacterial resistance mechanism that provides novel insights into the biology of bacterial secretion. |
| Databáze: | OpenAIRE |
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