The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1099/mic.0.000767 http://hdl.handle.net/11449/188862 |
Resumo: | Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene. |
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The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levelsCopper resistanceGene expressionShort upstream ORFXanthomonas citriUpstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Departamento de Bioquímica e Tecnologia Química Instituto de Química UNESP Universidade Estadual PaulistaDepartamento de Bioprocessos e Biotecnologia Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista UNESPDepartamento de Bioquímica e Microbiologia Instituto de Biociências UNESP Universidade Estadual PaulistaDepartment of Biochemistry University of CaliforniaDepartamento de Bioquímica e Tecnologia Química Instituto de Química UNESP Universidade Estadual PaulistaDepartamento de Bioprocessos e Biotecnologia Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista UNESPDepartamento de Bioquímica e Microbiologia Instituto de Biociências UNESP Universidade Estadual PaulistaFAPESP: 2005/04036-3Universidade Estadual Paulista (Unesp)University of CaliforniaDe Freitas, Eliane Cristina [UNESP]Ucci, Amanda Piovesan [UNESP]Teixeira, Elaine Cristina [UNESP]Pedroso, Gisele Audrei [UNESP]Hilario, Eduardo [UNESP]Bertolazzi Zocca, Vitória Fernanda [UNESP]De Paiva, Gabriela Barbosa [UNESP]Ferreira, Henrique [UNESP]Pedrolli, Danielle Biscaro [UNESP]Bertolini, Maria Célia [UNESP]2019-10-06T16:21:38Z2019-10-06T16:21:38Z2019-03-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article355-365http://dx.doi.org/10.1099/mic.0.000767Microbiology (United Kingdom), v. 165, n. 3, p. 355-365, 2019.1465-20801350-0872http://hdl.handle.net/11449/18886210.1099/mic.0.0007672-s2.0-850632271078817669953838863Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMicrobiology (United Kingdom)info:eu-repo/semantics/openAccess2021-10-23T12:39:39Zoai:repositorio.unesp.br:11449/188862Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T12:39:39Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
title |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
spellingShingle |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels De Freitas, Eliane Cristina [UNESP] Copper resistance Gene expression Short upstream ORF Xanthomonas citri |
title_short |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
title_full |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
title_fullStr |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
title_full_unstemmed |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
title_sort |
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels |
author |
De Freitas, Eliane Cristina [UNESP] |
author_facet |
De Freitas, Eliane Cristina [UNESP] Ucci, Amanda Piovesan [UNESP] Teixeira, Elaine Cristina [UNESP] Pedroso, Gisele Audrei [UNESP] Hilario, Eduardo [UNESP] Bertolazzi Zocca, Vitória Fernanda [UNESP] De Paiva, Gabriela Barbosa [UNESP] Ferreira, Henrique [UNESP] Pedrolli, Danielle Biscaro [UNESP] Bertolini, Maria Célia [UNESP] |
author_role |
author |
author2 |
Ucci, Amanda Piovesan [UNESP] Teixeira, Elaine Cristina [UNESP] Pedroso, Gisele Audrei [UNESP] Hilario, Eduardo [UNESP] Bertolazzi Zocca, Vitória Fernanda [UNESP] De Paiva, Gabriela Barbosa [UNESP] Ferreira, Henrique [UNESP] Pedrolli, Danielle Biscaro [UNESP] Bertolini, Maria Célia [UNESP] |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) University of California |
dc.contributor.author.fl_str_mv |
De Freitas, Eliane Cristina [UNESP] Ucci, Amanda Piovesan [UNESP] Teixeira, Elaine Cristina [UNESP] Pedroso, Gisele Audrei [UNESP] Hilario, Eduardo [UNESP] Bertolazzi Zocca, Vitória Fernanda [UNESP] De Paiva, Gabriela Barbosa [UNESP] Ferreira, Henrique [UNESP] Pedrolli, Danielle Biscaro [UNESP] Bertolini, Maria Célia [UNESP] |
dc.subject.por.fl_str_mv |
Copper resistance Gene expression Short upstream ORF Xanthomonas citri |
topic |
Copper resistance Gene expression Short upstream ORF Xanthomonas citri |
description |
Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-10-06T16:21:38Z 2019-10-06T16:21:38Z 2019-03-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1099/mic.0.000767 Microbiology (United Kingdom), v. 165, n. 3, p. 355-365, 2019. 1465-2080 1350-0872 http://hdl.handle.net/11449/188862 10.1099/mic.0.000767 2-s2.0-85063227107 8817669953838863 |
url |
http://dx.doi.org/10.1099/mic.0.000767 http://hdl.handle.net/11449/188862 |
identifier_str_mv |
Microbiology (United Kingdom), v. 165, n. 3, p. 355-365, 2019. 1465-2080 1350-0872 10.1099/mic.0.000767 2-s2.0-85063227107 8817669953838863 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Microbiology (United Kingdom) |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
355-365 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1797789993814458368 |