A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase
ABSTRACT 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...
Guardado en:
| Autores principales: | , , , , , , , , , , , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
American Society for Microbiology
2016
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/3b9f916136964f4eb118797ed067be1e |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:3b9f916136964f4eb118797ed067be1e |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:3b9f916136964f4eb118797ed067be1e2021-11-15T15:50:14ZA Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase10.1128/mBio.00412-162150-7511https://doaj.org/article/3b9f916136964f4eb118797ed067be1e2016-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00412-16https://doaj.org/toc/2150-7511ABSTRACT 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.J. Hiroshi MorisakiPeter A. SmithShailesh V. DateKimberly K. KajiharaChau Linda TruongZora ModrusanDonghong YanJing KangMin XuIshita M. ShahRobert MintzerEric M. KofoedTommy K. CheungDavid ArnottMichael F. T. KoehlerChristopher E. HeiseEric J. BrownMan-Wah TanWouter L. W. HazenbosAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 5 (2016) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Microbiology QR1-502 |
| spellingShingle |
Microbiology QR1-502 J. Hiroshi Morisaki Peter A. Smith Shailesh V. Date Kimberly K. Kajihara Chau Linda Truong Zora Modrusan Donghong Yan Jing Kang Min Xu Ishita M. Shah Robert Mintzer Eric M. Kofoed Tommy K. Cheung David Arnott Michael F. T. Koehler Christopher E. Heise Eric J. Brown Man-Wah Tan Wouter L. W. Hazenbos A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| description |
ABSTRACT 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. |
| format |
article |
| author |
J. Hiroshi Morisaki Peter A. Smith Shailesh V. Date Kimberly K. Kajihara Chau Linda Truong Zora Modrusan Donghong Yan Jing Kang Min Xu Ishita M. Shah Robert Mintzer Eric M. Kofoed Tommy K. Cheung David Arnott Michael F. T. Koehler Christopher E. Heise Eric J. Brown Man-Wah Tan Wouter L. W. Hazenbos |
| author_facet |
J. Hiroshi Morisaki Peter A. Smith Shailesh V. Date Kimberly K. Kajihara Chau Linda Truong Zora Modrusan Donghong Yan Jing Kang Min Xu Ishita M. Shah Robert Mintzer Eric M. Kofoed Tommy K. Cheung David Arnott Michael F. T. Koehler Christopher E. Heise Eric J. Brown Man-Wah Tan Wouter L. W. Hazenbos |
| author_sort |
J. Hiroshi Morisaki |
| title |
A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| title_short |
A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| title_full |
A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| title_fullStr |
A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| title_full_unstemmed |
A Putative Bacterial ABC Transporter Circumvents the Essentiality of Signal Peptidase |
| title_sort |
putative bacterial abc transporter circumvents the essentiality of signal peptidase |
| publisher |
American Society for Microbiology |
| publishDate |
2016 |
| url |
https://doaj.org/article/3b9f916136964f4eb118797ed067be1e |
| work_keys_str_mv |
AT jhiroshimorisaki aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT peterasmith aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT shaileshvdate aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT kimberlykkajihara aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT chaulindatruong aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT zoramodrusan aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT donghongyan aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT jingkang aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT minxu aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ishitamshah aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT robertmintzer aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ericmkofoed aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT tommykcheung aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT davidarnott aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT michaelftkoehler aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT christophereheise aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ericjbrown aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT manwahtan aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT wouterlwhazenbos aputativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT jhiroshimorisaki putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT peterasmith putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT shaileshvdate putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT kimberlykkajihara putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT chaulindatruong putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT zoramodrusan putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT donghongyan putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT jingkang putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT minxu putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ishitamshah putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT robertmintzer putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ericmkofoed putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT tommykcheung putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT davidarnott putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT michaelftkoehler putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT christophereheise putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT ericjbrown putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT manwahtan putativebacterialabctransportercircumventstheessentialityofsignalpeptidase AT wouterlwhazenbos putativebacterialabctransportercircumventstheessentialityofsignalpeptidase |
| _version_ |
1718427452917678080 |