Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein

ABSTRACT Burkholderia thailandensis produces a family of polyketide-peptide molecules called bactobolins, some of which are potent antibiotics. We found that growth of B. thailandensis at 30°C versus that at 37°C resulted in increased production of bactobolins. We purified the three most abundant ba...

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Autores principales:	Josephine R. Chandler, Thao T. Truong, Patricia M. Silva, Mohammad R. Seyedsayamdost, Gavin Carr, Matthew Radey, Michael A. Jacobs, Elizabeth H. Sims, Jon Clardy, E. Peter Greenberg
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/fd866855a199477a85816d05ef1e7055
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spelling	oai:doaj.org-article:fd866855a199477a85816d05ef1e70552021-11-15T15:39:11ZBactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein10.1128/mBio.00499-122150-7511https://doaj.org/article/fd866855a199477a85816d05ef1e70552012-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00499-12https://doaj.org/toc/2150-7511ABSTRACT Burkholderia thailandensis produces a family of polyketide-peptide molecules called bactobolins, some of which are potent antibiotics. We found that growth of B. thailandensis at 30°C versus that at 37°C resulted in increased production of bactobolins. We purified the three most abundant bactobolins and determined their activities against a battery of bacteria and mouse fibroblasts. Two of the three compounds showed strong activities against both bacteria and fibroblasts. The third analog was much less potent in both assays. These results suggested that the target of bactobolins might be conserved across bacteria and mammalian cells. To learn about the mechanism of bactobolin activity, we isolated four spontaneous bactobolin-resistant Bacillus subtilis mutants. We used genomic sequencing technology to show that each of the four resistant variants had mutations in rplB, which codes for the 50S ribosome-associated L2 protein. Ectopic expression of a mutant rplB gene in wild-type B. subtilis conferred bactobolin resistance. Finally, the L2 mutations did not confer resistance to other antibiotics known to interfere with ribosome function. Our data indicate that bactobolins target the L2 protein or a nearby site and that this is not the target of other antibiotics. We presume that the mammalian target of bactobolins involves the eukaryotic homolog of L2 (L8e). IMPORTANCE Currently available antibiotics target surprisingly few cellular functions, and there is a need to identify novel antibiotic targets. We have been interested in the Burkholderia thailandensis bactobolins, and we sought to learn about the target of bactobolin activity by mapping spontaneous resistance mutations in the bactobolin-sensitive Bacillus subtilis. Our results indicate that the bactobolin target is the 50S ribosome-associated L2 protein or a region of the ribosome affected by L2. Bactobolin-resistant mutants are not resistant to other known ribosome inhibitors. Our evidence indicates that bactobolins interact with a novel antibiotic target.Josephine R. ChandlerThao T. TruongPatricia M. SilvaMohammad R. SeyedsayamdostGavin CarrMatthew RadeyMichael A. JacobsElizabeth H. SimsJon ClardyE. Peter GreenbergAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 6 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Josephine R. Chandler Thao T. Truong Patricia M. Silva Mohammad R. Seyedsayamdost Gavin Carr Matthew Radey Michael A. Jacobs Elizabeth H. Sims Jon Clardy E. Peter Greenberg Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
description	ABSTRACT Burkholderia thailandensis produces a family of polyketide-peptide molecules called bactobolins, some of which are potent antibiotics. We found that growth of B. thailandensis at 30°C versus that at 37°C resulted in increased production of bactobolins. We purified the three most abundant bactobolins and determined their activities against a battery of bacteria and mouse fibroblasts. Two of the three compounds showed strong activities against both bacteria and fibroblasts. The third analog was much less potent in both assays. These results suggested that the target of bactobolins might be conserved across bacteria and mammalian cells. To learn about the mechanism of bactobolin activity, we isolated four spontaneous bactobolin-resistant Bacillus subtilis mutants. We used genomic sequencing technology to show that each of the four resistant variants had mutations in rplB, which codes for the 50S ribosome-associated L2 protein. Ectopic expression of a mutant rplB gene in wild-type B. subtilis conferred bactobolin resistance. Finally, the L2 mutations did not confer resistance to other antibiotics known to interfere with ribosome function. Our data indicate that bactobolins target the L2 protein or a nearby site and that this is not the target of other antibiotics. We presume that the mammalian target of bactobolins involves the eukaryotic homolog of L2 (L8e). IMPORTANCE Currently available antibiotics target surprisingly few cellular functions, and there is a need to identify novel antibiotic targets. We have been interested in the Burkholderia thailandensis bactobolins, and we sought to learn about the target of bactobolin activity by mapping spontaneous resistance mutations in the bactobolin-sensitive Bacillus subtilis. Our results indicate that the bactobolin target is the 50S ribosome-associated L2 protein or a region of the ribosome affected by L2. Bactobolin-resistant mutants are not resistant to other known ribosome inhibitors. Our evidence indicates that bactobolins interact with a novel antibiotic target.
format	article
author	Josephine R. Chandler Thao T. Truong Patricia M. Silva Mohammad R. Seyedsayamdost Gavin Carr Matthew Radey Michael A. Jacobs Elizabeth H. Sims Jon Clardy E. Peter Greenberg
author_facet	Josephine R. Chandler Thao T. Truong Patricia M. Silva Mohammad R. Seyedsayamdost Gavin Carr Matthew Radey Michael A. Jacobs Elizabeth H. Sims Jon Clardy E. Peter Greenberg
author_sort	Josephine R. Chandler
title	Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
title_short	Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
title_full	Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
title_fullStr	Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
title_full_unstemmed	Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein
title_sort	bactobolin resistance is conferred by mutations in the l2 ribosomal protein
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/fd866855a199477a85816d05ef1e7055
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Bactobolin Resistance Is Conferred by Mutations in the L2 Ribosomal Protein

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