Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like

Detalhes bibliográficos
Autor(a) principal: Silva, Lucas Araújo Castro e
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/2470
Resumo: The PAM2 motif is largely distributed in eukaryotic proteins and was initially identified as a binding site for poly (A) binding proteins (PABP), which play a role in RNA metabolism. Despite the PAM2 motif relevance, a comprehensive analysis of its occurrence in proteins of the plant kingdom is still absent. In the present study, the PAM2-like protein family was phylogenetically characterized in Arabidopsis, soybean and rice. Totally, 19 PAM2-like sequences were identified in the Arabidopsis genome, while 31 members of the family were found in soybean and 22 members in rice. The PAM2-like family was subdivided in 7 subfamilies (A-G) based on the conservation of the sequences and in the structure of the motifs. Among them, the ERD15-like subfamily, whose name derives from the ERD15 (Early Responsive to Dehydration 15) Arabidopsis protein, originally identified by its rapid induction in response to dehydration, was characterized extensively, concerning its phylogeny, dehydration responsiveness and cellular function. Microarray data from stressed plants shows that both members of the ERD15-like subfamily in Arabidopsis, AtERD15 (AT2G41430) e AtERL15 (AT4G14270), are responsive to drought. The drought response profile of 4 of the 6 soybean ERD15-like subfamily members was also investigated. It was observed that the pair Glyma14g05980 and Glyma02g42860 is continuously induced as the stress progress, whereas the pair Glyma11g20940 and Glyma04g28560 is initially induced, but has a reduction on its expression level when the stress becomes more severe, indicating a possible functional divergence in the ERD15-like subfamily. The superexpression and the suppression of the AtERD15 gene were evaluated regarding to sensibility to ABA and tolerance to drought. A correlation between AtERD15 (AT2G41430) expression level and the germination rate of seeds in the presence of the phytohormone ABA was observed. Opposingly, a negative correlation was observed for the level of AtERD15 transcript and the root growth in plants stressed by PEG, as well as for the drought stress tolerance. The soybean, GmERD15-like (Glyma02g42860), and rice, OsERD15-like (Os07g46670), homologs shown partial phenotype complementation for ABA susceptibility during germination, as presented by AtERD15, but, on the contrary, promoted increased drought tolerance on stressed plants. Collectively, these results imply that the members of the ERD15-like family diverge in function. Under normal conditions, the proteins AtERD15, GmERD15-like and OsERD15-like are localized in the cytoplasm. Nonetheless, under osmotic stress, the subcellular localization profile of these proteins is modified, as they increase their concentration around the nucleus. The OsERD15-like protein (Os07g46670) was translocated to the nucleus under osmotic stress conditions. Under transient expression in tobacco leaves, the three proteins exhibited the same localization pattern, being present in the cytoplasm as well as in the nucleus, except in the nucleolus. Finally, it was demonstrated that the protein GmERD15-like (Glyma02g42860) is capable of binding to the promoters of the genes CDC2 (Glyma06g17640) and F-Box (Glyma13g43730) and that the superexpression of GmERD15-like in soybean protoplasts led the induction of both genes, in addition to DNAJ (Glyma14g31810). CDC2, DNAJ and F-Box are all induced by drought. These results indicate that GmERD15-like controls a selected group of genes induced by hydric stress.
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spelling Silva, Lucas Araújo Castro ehttp://lattes.cnpq.br/6413364405549031Pereira, Welison Andradehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4742902Y6Bevitori, Rosângelahttp://lattes.cnpq.br/3979573959730493Fontes, Elizabeth Pacheco Batistahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781848H2Deguchi, Michihitohttp://lattes.cnpq.br/95064003330225642015-03-26T13:07:38Z2014-11-282015-03-26T13:07:38Z2014-02-28SILVA, Lucas Araújo Castro e. Analysis of the PAM2-like protein family in soybean: functional divergence in the ERD15-like subfamily. 2014. 101 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2014.http://locus.ufv.br/handle/123456789/2470The PAM2 motif is largely distributed in eukaryotic proteins and was initially identified as a binding site for poly (A) binding proteins (PABP), which play a role in RNA metabolism. Despite the PAM2 motif relevance, a comprehensive analysis of its occurrence in proteins of the plant kingdom is still absent. In the present study, the PAM2-like protein family was phylogenetically characterized in Arabidopsis, soybean and rice. Totally, 19 PAM2-like sequences were identified in the Arabidopsis genome, while 31 members of the family were found in soybean and 22 members in rice. The PAM2-like family was subdivided in 7 subfamilies (A-G) based on the conservation of the sequences and in the structure of the motifs. Among them, the ERD15-like subfamily, whose name derives from the ERD15 (Early Responsive to Dehydration 15) Arabidopsis protein, originally identified by its rapid induction in response to dehydration, was characterized extensively, concerning its phylogeny, dehydration responsiveness and cellular function. Microarray data from stressed plants shows that both members of the ERD15-like subfamily in Arabidopsis, AtERD15 (AT2G41430) e AtERL15 (AT4G14270), are responsive to drought. The drought response profile of 4 of the 6 soybean ERD15-like subfamily members was also investigated. It was observed that the pair Glyma14g05980 and Glyma02g42860 is continuously induced as the stress progress, whereas the pair Glyma11g20940 and Glyma04g28560 is initially induced, but has a reduction on its expression level when the stress becomes more severe, indicating a possible functional divergence in the ERD15-like subfamily. The superexpression and the suppression of the AtERD15 gene were evaluated regarding to sensibility to ABA and tolerance to drought. A correlation between AtERD15 (AT2G41430) expression level and the germination rate of seeds in the presence of the phytohormone ABA was observed. Opposingly, a negative correlation was observed for the level of AtERD15 transcript and the root growth in plants stressed by PEG, as well as for the drought stress tolerance. The soybean, GmERD15-like (Glyma02g42860), and rice, OsERD15-like (Os07g46670), homologs shown partial phenotype complementation for ABA susceptibility during germination, as presented by AtERD15, but, on the contrary, promoted increased drought tolerance on stressed plants. Collectively, these results imply that the members of the ERD15-like family diverge in function. Under normal conditions, the proteins AtERD15, GmERD15-like and OsERD15-like are localized in the cytoplasm. Nonetheless, under osmotic stress, the subcellular localization profile of these proteins is modified, as they increase their concentration around the nucleus. The OsERD15-like protein (Os07g46670) was translocated to the nucleus under osmotic stress conditions. Under transient expression in tobacco leaves, the three proteins exhibited the same localization pattern, being present in the cytoplasm as well as in the nucleus, except in the nucleolus. Finally, it was demonstrated that the protein GmERD15-like (Glyma02g42860) is capable of binding to the promoters of the genes CDC2 (Glyma06g17640) and F-Box (Glyma13g43730) and that the superexpression of GmERD15-like in soybean protoplasts led the induction of both genes, in addition to DNAJ (Glyma14g31810). CDC2, DNAJ and F-Box are all induced by drought. These results indicate that GmERD15-like controls a selected group of genes induced by hydric stress.O motivo PAM2 está largamente distribuído em proteínas eucarióticas e foi incialmente identificado como um sítio de ligação a proteínas que se ligam à cauda poli (A) (PABP), que participam do metabolismo de RNA. Apesar da relevância do motivo PAM2, uma análise compreensiva de sua ocorrência em proteínas do reino vegetal ainda não foi conduzida. Nesta investigação, a família das proteínas PAM2-like foi caracterizada filogeneticamente em Arabidopsis, soja e arroz. Um total de 19 sequências PAM2-like foi identificado no genoma de Arabidopsis, enquanto que em soja foram encontrados 31 membros e em arroz 22 membros da família. A família PAM2-like foi subdividida em 7 subfamílias (A-G) baseado na conservação de sequências e na estrutura de motivos característicos. Entre elas, a subfamília ERD15-like, cujo nome foi derivado da proteína ERD15 (Early Responsive to Dehydration 15) de Arabidopsis, identificada originalmente por sua rápida indução em resposta à desidratação, foi caracterizada extensivamente, em relação à sua filogenia, responsividade à seca e função celular. Foi observado em dados de micro arranjo de plantas estressadas que os dois membros da subfamília ERD15-like de Arabidopsis, AtERD15 (AT2G41430) e AtERL15 (AT4G14270), são responsivos à seca. O perfil de resposta ao estresse causado pela seca de 4 dos 6 membros da subfamília de soja também foi avaliado, sendo que o par Glyma14g05980 e Glyma02g42860 progressão do estresse, ao é continuamente induzido com a passo que o par Glyma11g20940 e Glyma04g28560 é inicialmente induzido, mas tem seu nível de expressão reduzido no estresse mais severo, indicando uma possível divergência funcional na subfamília ERD15-like. A superexpressão e supressão do gene AtERD15 foram avaliadas quanto à sensibilidade à ABA e tolerância à seca. Foi observada uma correlação entre o nível de expressão de AtERD15 (AT2G41430) e a capacidade de germinação de sementes na presença do fito hormônio ABA. Contrariamente, uma correlação negativa foi observada entre o nível de transcrito de AtERD15 e o crescimento de raiz em plantas estressadas por PEG, assim como a tolerância a estresse causado pela seca. Os homólogos da soja, GmERD15-like (Glyma02g42860), e de arroz, OsERD15- like (Os07g46670) complementaram parcialmente o fenótipo de suscetibilidade à ABA durante a germinação apresentado por AtERD15, mas ao contrário de AtERD15, promoveram um aumento da tolerância à seca em plantas estressadas. Coletivamente, estes resultados implicam que os membros da família ERD15 divergem funcionalmente. As proteínas AtERD15, GmERD15- like e OsERD15-like são localizadas no citoplasma em condições normais. Entretanto, em resposta a estresse osmótico o perfil de localização das proteínas é modificado, sendo concentradas ao redor do núcleo. A proteína OsERD15-like (Os07g46670) foi translocada para o núcleo em condições de estresse osmótico. Em expressão transiente em folhas de tabaco, todas três proteínas exibiram o mesmo padrão de localização, estando presentes tanto no citoplasma, como no núcleo, com exclusão do nucléolo. Por fim, foi demonstrado que a proteína GmERD15-like (Glyma02g42860) tem a capacidade de ligação aos promotores dos genes CDC2 (Glyma06g17460) e F- Box (Glyma13g43730) e que sua superexpressão de GmERD15-like em protoplastos de soja levou a indução da expressão de ambos, além do gene DNAJ (Glyma14g31810). Tanto CDC2, como DNAJ e F-Box são induzidos por seca. Estes resultados indicam que GmERD15-like controla um conjunto seleto de genes induzidos por estresse hídrico.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em Bioquímica AgrícolaUFVBRBioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animalProteínas - AnálisesSojaArrozERD15PAM2Plantas - Estresse hídricoProteins - analysisSoybeansRiceERD15PAM2Plants - Water stressCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULARAnálise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-likeAnalysis of the PAM2-like protein family in soybean: functional divergence in the ERD15-like subfamilyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf2486616https://locus.ufv.br//bitstream/123456789/2470/1/texto%20completo.pdf8e71d34d179d35f9d49ed3fc02fdd5f5MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain251351https://locus.ufv.br//bitstream/123456789/2470/2/texto%20completo.pdf.txt364e055ac96d8a1036ef118a6137a391MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3583https://locus.ufv.br//bitstream/123456789/2470/3/texto%20completo.pdf.jpg1b6d88d9c7f4623728aa196ae051be00MD53123456789/24702016-04-08 23:05:39.938oai:locus.ufv.br:123456789/2470Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:05:39LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
dc.title.alternative.eng.fl_str_mv Analysis of the PAM2-like protein family in soybean: functional divergence in the ERD15-like subfamily
title Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
spellingShingle Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
Silva, Lucas Araújo Castro e
Proteínas - Análises
Soja
Arroz
ERD15
PAM2
Plantas - Estresse hídrico
Proteins - analysis
Soybeans
Rice
ERD15
PAM2
Plants - Water stress
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
title_short Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
title_full Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
title_fullStr Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
title_full_unstemmed Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
title_sort Análise da família das proteínas PAM2-like em soja: divergência funcional da subfamília das ERD15-like
author Silva, Lucas Araújo Castro e
author_facet Silva, Lucas Araújo Castro e
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/6413364405549031
dc.contributor.author.fl_str_mv Silva, Lucas Araújo Castro e
dc.contributor.advisor-co1.fl_str_mv Pereira, Welison Andrade
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4742902Y6
dc.contributor.advisor-co2.fl_str_mv Bevitori, Rosângela
dc.contributor.advisor-co2Lattes.fl_str_mv http://lattes.cnpq.br/3979573959730493
dc.contributor.advisor1.fl_str_mv Fontes, Elizabeth Pacheco Batista
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781848H2
dc.contributor.referee1.fl_str_mv Deguchi, Michihito
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/9506400333022564
contributor_str_mv Pereira, Welison Andrade
Bevitori, Rosângela
Fontes, Elizabeth Pacheco Batista
Deguchi, Michihito
dc.subject.por.fl_str_mv Proteínas - Análises
Soja
Arroz
ERD15
PAM2
Plantas - Estresse hídrico
topic Proteínas - Análises
Soja
Arroz
ERD15
PAM2
Plantas - Estresse hídrico
Proteins - analysis
Soybeans
Rice
ERD15
PAM2
Plants - Water stress
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
dc.subject.eng.fl_str_mv Proteins - analysis
Soybeans
Rice
ERD15
PAM2
Plants - Water stress
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
description The PAM2 motif is largely distributed in eukaryotic proteins and was initially identified as a binding site for poly (A) binding proteins (PABP), which play a role in RNA metabolism. Despite the PAM2 motif relevance, a comprehensive analysis of its occurrence in proteins of the plant kingdom is still absent. In the present study, the PAM2-like protein family was phylogenetically characterized in Arabidopsis, soybean and rice. Totally, 19 PAM2-like sequences were identified in the Arabidopsis genome, while 31 members of the family were found in soybean and 22 members in rice. The PAM2-like family was subdivided in 7 subfamilies (A-G) based on the conservation of the sequences and in the structure of the motifs. Among them, the ERD15-like subfamily, whose name derives from the ERD15 (Early Responsive to Dehydration 15) Arabidopsis protein, originally identified by its rapid induction in response to dehydration, was characterized extensively, concerning its phylogeny, dehydration responsiveness and cellular function. Microarray data from stressed plants shows that both members of the ERD15-like subfamily in Arabidopsis, AtERD15 (AT2G41430) e AtERL15 (AT4G14270), are responsive to drought. The drought response profile of 4 of the 6 soybean ERD15-like subfamily members was also investigated. It was observed that the pair Glyma14g05980 and Glyma02g42860 is continuously induced as the stress progress, whereas the pair Glyma11g20940 and Glyma04g28560 is initially induced, but has a reduction on its expression level when the stress becomes more severe, indicating a possible functional divergence in the ERD15-like subfamily. The superexpression and the suppression of the AtERD15 gene were evaluated regarding to sensibility to ABA and tolerance to drought. A correlation between AtERD15 (AT2G41430) expression level and the germination rate of seeds in the presence of the phytohormone ABA was observed. Opposingly, a negative correlation was observed for the level of AtERD15 transcript and the root growth in plants stressed by PEG, as well as for the drought stress tolerance. The soybean, GmERD15-like (Glyma02g42860), and rice, OsERD15-like (Os07g46670), homologs shown partial phenotype complementation for ABA susceptibility during germination, as presented by AtERD15, but, on the contrary, promoted increased drought tolerance on stressed plants. Collectively, these results imply that the members of the ERD15-like family diverge in function. Under normal conditions, the proteins AtERD15, GmERD15-like and OsERD15-like are localized in the cytoplasm. Nonetheless, under osmotic stress, the subcellular localization profile of these proteins is modified, as they increase their concentration around the nucleus. The OsERD15-like protein (Os07g46670) was translocated to the nucleus under osmotic stress conditions. Under transient expression in tobacco leaves, the three proteins exhibited the same localization pattern, being present in the cytoplasm as well as in the nucleus, except in the nucleolus. Finally, it was demonstrated that the protein GmERD15-like (Glyma02g42860) is capable of binding to the promoters of the genes CDC2 (Glyma06g17640) and F-Box (Glyma13g43730) and that the superexpression of GmERD15-like in soybean protoplasts led the induction of both genes, in addition to DNAJ (Glyma14g31810). CDC2, DNAJ and F-Box are all induced by drought. These results indicate that GmERD15-like controls a selected group of genes induced by hydric stress.
publishDate 2014
dc.date.available.fl_str_mv 2014-11-28
2015-03-26T13:07:38Z
dc.date.issued.fl_str_mv 2014-02-28
dc.date.accessioned.fl_str_mv 2015-03-26T13:07:38Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv SILVA, Lucas Araújo Castro e. Analysis of the PAM2-like protein family in soybean: functional divergence in the ERD15-like subfamily. 2014. 101 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2014.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/2470
identifier_str_mv SILVA, Lucas Araújo Castro e. Analysis of the PAM2-like protein family in soybean: functional divergence in the ERD15-like subfamily. 2014. 101 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2014.
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