Cargando…
Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR
Stenotrophomonas maltophilia contains an operon comprising mfsB and mfsC, which encode membrane transporters in the major facilitator superfamily (MFS). The results of the topological analysis predicted that both MfsB and MfsC possess 12 transmembrane helices with the N- and C-termini located inside...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9342713/ https://www.ncbi.nlm.nih.gov/pubmed/35913917 http://dx.doi.org/10.1371/journal.pone.0272388 |
_version_ | 1784760883248889856 |
---|---|
author | Boonyakanog, Angkana Charoenlap, Nisanart Chattrakarn, Sorayut Vattanaviboon, Paiboon Mongkolsuk, Skorn |
author_facet | Boonyakanog, Angkana Charoenlap, Nisanart Chattrakarn, Sorayut Vattanaviboon, Paiboon Mongkolsuk, Skorn |
author_sort | Boonyakanog, Angkana |
collection | PubMed |
description | Stenotrophomonas maltophilia contains an operon comprising mfsB and mfsC, which encode membrane transporters in the major facilitator superfamily (MFS). The results of the topological analysis predicted that both MfsB and MfsC possess 12 transmembrane helices with the N- and C-termini located inside the cells. The deletion of mfsC increased the susceptibility to diamide, a chemical oxidizing agent, but not to antibiotics and oxidative stress-generating substances relative to wild-type K279a. Moreover, no altered phenotype was observed against all tested substances for the ΔmfsB mutant. The results of the expression analysis revealed that the mfsBC expression was significantly induced by exposure to diamide. The diamide-induced gene expression was mediated by DitR, a TetR-type transcriptional regulator encoded by smlt0547. A constitutively high expression of mfsC in the ditR mutant indicated that DitR acts as a transcriptional repressor of mfsBC under physiological conditions. Purified DitR was bound to three sites spanning from position + 21 to -57, corresponding to the putative mfsBC promoter sequence, thereby interfering with the binding of RNA polymerase. The results of electrophoretic mobility shift assays illustrated that the treatment of purified DitR with diamide caused the release of DitR from the mfsBC promoter region, and the diamide sensing mechanism of DitR required two conserved cysteine residues, Cys92 and Cys127. This suggests that exposure to diamide can oxidize DitR through the oxidation of cysteine residues, leading to its release from the promoter, thus allowing mfsBC transcription. Overall, MfsC and DitR play a role in adaptive resistance against the diamide of S. maltophilia. |
format | Online Article Text |
id | pubmed-9342713 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-93427132022-08-02 Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR Boonyakanog, Angkana Charoenlap, Nisanart Chattrakarn, Sorayut Vattanaviboon, Paiboon Mongkolsuk, Skorn PLoS One Research Article Stenotrophomonas maltophilia contains an operon comprising mfsB and mfsC, which encode membrane transporters in the major facilitator superfamily (MFS). The results of the topological analysis predicted that both MfsB and MfsC possess 12 transmembrane helices with the N- and C-termini located inside the cells. The deletion of mfsC increased the susceptibility to diamide, a chemical oxidizing agent, but not to antibiotics and oxidative stress-generating substances relative to wild-type K279a. Moreover, no altered phenotype was observed against all tested substances for the ΔmfsB mutant. The results of the expression analysis revealed that the mfsBC expression was significantly induced by exposure to diamide. The diamide-induced gene expression was mediated by DitR, a TetR-type transcriptional regulator encoded by smlt0547. A constitutively high expression of mfsC in the ditR mutant indicated that DitR acts as a transcriptional repressor of mfsBC under physiological conditions. Purified DitR was bound to three sites spanning from position + 21 to -57, corresponding to the putative mfsBC promoter sequence, thereby interfering with the binding of RNA polymerase. The results of electrophoretic mobility shift assays illustrated that the treatment of purified DitR with diamide caused the release of DitR from the mfsBC promoter region, and the diamide sensing mechanism of DitR required two conserved cysteine residues, Cys92 and Cys127. This suggests that exposure to diamide can oxidize DitR through the oxidation of cysteine residues, leading to its release from the promoter, thus allowing mfsBC transcription. Overall, MfsC and DitR play a role in adaptive resistance against the diamide of S. maltophilia. Public Library of Science 2022-08-01 /pmc/articles/PMC9342713/ /pubmed/35913917 http://dx.doi.org/10.1371/journal.pone.0272388 Text en © 2022 Boonyakanog et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Boonyakanog, Angkana Charoenlap, Nisanart Chattrakarn, Sorayut Vattanaviboon, Paiboon Mongkolsuk, Skorn Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title | Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title_full | Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title_fullStr | Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title_full_unstemmed | Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title_short | Contribution of Stenotrophomonas maltophilia MfsC transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor DitR |
title_sort | contribution of stenotrophomonas maltophilia mfsc transporter to protection against diamide and the regulation of its expression by the diamide responsive repressor ditr |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9342713/ https://www.ncbi.nlm.nih.gov/pubmed/35913917 http://dx.doi.org/10.1371/journal.pone.0272388 |
work_keys_str_mv | AT boonyakanogangkana contributionofstenotrophomonasmaltophiliamfsctransportertoprotectionagainstdiamideandtheregulationofitsexpressionbythediamideresponsiverepressorditr AT charoenlapnisanart contributionofstenotrophomonasmaltophiliamfsctransportertoprotectionagainstdiamideandtheregulationofitsexpressionbythediamideresponsiverepressorditr AT chattrakarnsorayut contributionofstenotrophomonasmaltophiliamfsctransportertoprotectionagainstdiamideandtheregulationofitsexpressionbythediamideresponsiverepressorditr AT vattanaviboonpaiboon contributionofstenotrophomonasmaltophiliamfsctransportertoprotectionagainstdiamideandtheregulationofitsexpressionbythediamideresponsiverepressorditr AT mongkolsukskorn contributionofstenotrophomonasmaltophiliamfsctransportertoprotectionagainstdiamideandtheregulationofitsexpressionbythediamideresponsiverepressorditr |