Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa

Detalhes bibliográficos
Autor(a) principal: Fonseca, Erica L.
Data de Publicação: 2015
Outros Autores: Vicente, Ana Carolina Paulo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/12550
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genética Molecular de Microrganismos. Rio de Janeiro, RJ, Brasil.
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spelling Fonseca, Erica L.Vicente, Ana Carolina Paulo2016-01-19T10:40:12Z2016-01-19T10:40:12Z2015FONSECA, Érica L.; VICENTE, Ana Carolina Paulo. Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa. F1000Research, 17p, 2015.10.12688/f1000research.2-99.v1https://www.arca.fiocruz.br/handle/icict/125502046-1402engF1000ResearchPolycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genética Molecular de Microrganismos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genética Molecular de Microrganismos. Rio de Janeiro, RJ, Brasil.The gene cassettes found in class 1 integrons are generally promoterless units composed by an open reading frame (ORF), a short 5' untranslated region (UTR) and a 3' recombination site ( attC). Fused gene cassettes are generated by partial or total loss of the attC from the first cassette in an array, creating, in some cases, a fusion with the ORF from the next cassette. These structures are rare and little is known about their mechanisms of mobilization and expression. The aim of this study was to evaluate the dynamic of mobilization and transcription of the gcu14-bla GES-1 /aacA4 gene cassette array, which harbours a fused gene cassette represented by bla GES-1 /aacA4. The cassette array was analyzed by Northern blot and real-time reverse transcription-polymerase chain reaction (RT-PCR) in order to assess the transcription mechanism of bla GES-1 /aacA4 fused cassette. Also, inverse polymerase chain reactions (PCR) were performed to detect the free circular forms of gcu14, bla GES-1 and aacA4. The Northern blot and real time RT-PCR revealed a polycistronic transcription, in which the fused cassette bla GES-1 /aacA4 is transcribed as a unique gene, while gcu14 (with a canonical attC recombination site) has a monocistronic transcription. The gcu14 cassette, closer to the weak configuration of cassette promoter (PcW), had a higher transcription level than bla GES-1/ aacA4, indicating that the cassette position affects the transcript amounts. The presence of ORF-11 at attI1, immediately preceding gcu14, and of a Shine-Dalgarno sequence upstream bla GES-1/ aacA4 composes a scenario for the occurrence of array translation. Inverse PCR generated amplicons corresponding to gcu14, gcu14-aacA4 and gcu14-bla GES-1/ aacA4 free circular forms, but not to bla GES-1 and aacA4 alone, indicating that the GES-1 truncated attC is not substrate of integrase activity and that these genes are mobilized together as a unique cassette. This study was original in showing the transcription of fused cassettes and in correlating cassette position with transcription.Clase 1 integronsFused gene cassetteGene cassetteTranscriptionPseudomonas aeruginosaPseudomonas aeruginosainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/12550/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALericafonseca_anavicente_IOC_2013.pdfericafonseca_anavicente_IOC_2013.pdfapplication/pdf1639856https://www.arca.fiocruz.br/bitstream/icict/12550/2/ericafonseca_anavicente_IOC_2013.pdf02ed4ceb6a1d7c9098f1e421c509f0d2MD52TEXTericafonseca_anavicente_IOC_2013.pdf.txtericafonseca_anavicente_IOC_2013.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
title Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
spellingShingle Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
Fonseca, Erica L.
Clase 1 integrons
Fused gene cassette
Gene cassette
Transcription
Pseudomonas aeruginosa
Pseudomonas aeruginosa
title_short Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
title_full Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
title_fullStr Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
title_full_unstemmed Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
title_sort Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa
author Fonseca, Erica L.
author_facet Fonseca, Erica L.
Vicente, Ana Carolina Paulo
author_role author
author2 Vicente, Ana Carolina Paulo
author2_role author
dc.contributor.author.fl_str_mv Fonseca, Erica L.
Vicente, Ana Carolina Paulo
dc.subject.en.pt_BR.fl_str_mv Clase 1 integrons
Fused gene cassette
Gene cassette
Transcription
Pseudomonas aeruginosa
topic Clase 1 integrons
Fused gene cassette
Gene cassette
Transcription
Pseudomonas aeruginosa
Pseudomonas aeruginosa
dc.subject.decs.pt_BR.fl_str_mv Pseudomonas aeruginosa
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genética Molecular de Microrganismos. Rio de Janeiro, RJ, Brasil.
publishDate 2015
dc.date.issued.fl_str_mv 2015
dc.date.accessioned.fl_str_mv 2016-01-19T10:40:12Z
dc.date.available.fl_str_mv 2016-01-19T10:40:12Z
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.citation.fl_str_mv FONSECA, Érica L.; VICENTE, Ana Carolina Paulo. Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa. F1000Research, 17p, 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/12550
dc.identifier.issn.none.fl_str_mv 10.12688/f1000research.2-99.v1
dc.identifier.eissn.none.fl_str_mv 2046-1402
identifier_str_mv FONSECA, Érica L.; VICENTE, Ana Carolina Paulo. Polycistronic transcription of fused cassettes and identification of translation initiation signals in an unusual gene cassette array from Pseudomonas aeruginosa. F1000Research, 17p, 2015.
10.12688/f1000research.2-99.v1
2046-1402
url https://www.arca.fiocruz.br/handle/icict/12550
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv F1000Research
publisher.none.fl_str_mv F1000Research
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/12550/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/12550/2/ericafonseca_anavicente_IOC_2013.pdf
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