Caracterização molecular e bioquímica de uma esterase macho-específica em Zaprionus indianus
Data
2008-02-21
Autores
Paulino, Rafael Marques [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Zaprionus indianus (Diptera:Drosophilidae) é uma espécie provavelmente de origem africana e que rapidamente se dispersou por parte do continente sul-americano. Hoje, indivíduos dessa espécie são encontrados numa amplitude latitudinal de 35°, do Uruguai a Belém (Brasil). Em Z. indianus e nos demais insetos, as esterases constituem um grupo multifuncional e heterogêneo de enzimas que participam da hidrólise de ésteres, além de estarem relacionadas a diversos processos metabólicos. Neste estudo, foram realizadas análises de esterases de Z. indianus e Drosophila melanogaster, em géis de poliacrilamida (PAGE) a 10% de concentração, em indivíduos dos dois sexos e em diferentes fases do desenvolvimento. Os resultados indicaram que algumas carboxilesterases (respectivamente EST-2 e EST-5 de Z. indianus e EST-6 D. melanogaster) apresentam atividade acentuada em machos. Estas enzimas mostraram similaridades nas duas espécies, tais como o mesmo padrão de inibição, expressão principalmente no estágio adulto e aumento da atividade enzimática com o aumento da idade dos indivíduos. As similaridades bioquímicas entre as enzimas dos dois drosofilídeos sugerem ortologia entre seus genes codificadores. Foram então realizadas reações de PCR, utilizando oligonucleotídeos iniciadores desenhados com base na seqüência do gene Est-6 de D. melanogaster e o DNA genômico de Z. indianus. O fragmento amplificado foi seqüenciado, com composição GC de 48,5% e similaridade de 73% com a seqüência do gene de Est-6 de D. melanogaster. A análise da razão das substituições sinônimas e não sinônimas sugere uma proteína sob ação de seleção normalizadora, onde as trocas sinônimas são em sua maioria neutras e as não sinônimas, na maioria das vezes deletérias, foram eliminadas pela seleção natural. A modelagem...
Zaprionus indianus has expanded its geographical distribution since its recent invasion of the South American continent. The first record data of only eight years, and the origin is probably the South Africa. Nowadays, this species can be found in a latitudinal range of 35º, from Uruguay to Belem (Brazil). Esterases comprise a multi-functional and heterogeneous group of enzymes that participated in ester hydrolysis. In insects, they are related to several metabolic processes, including the reproductive function. Esterase patterns in Z. indianus and Drosophila melanogaster were characterized in polyacrylamide gels (PAGE). Two - esterases from Z. indianus, EST-2 and EST-5, showed similarity preference for substrate - naftil acetate and patterns of inhibition as compared to the EST-6 of D. melanogaster, suggesting a possible role in reproductive biology for both enzymes. Biochemical characterization of these esterases and its differential expression in males, suggest orthology among their genes. In this work, the genomic DNA from Z. indianus was submitted to PCR experiments using 3 sets of D. melanogaster Est-6 sequence primers. The PCR products were directly sequenced; its GC content was 48.5% and presented a 73% similarity compared to Est-6 D. melanogaster sequence. Analysis of sequence data, by estimates of nonsynonymous/synonymous rate ratios, showed that the majority of sites evolves under strong or moderate negative selection (81%) and a minority of sites (19%). is under significant positive selection. Molecular modeling indicates that the fundamental structural features important for catalysis are conserved in the esterase of Z. indianus and human bilesalt activated lipase (BAL), which was used as structural model. Structural and sequence comparisons suggest the evolutionary relationship between ...(Complete abstract click electronic access below)
Zaprionus indianus has expanded its geographical distribution since its recent invasion of the South American continent. The first record data of only eight years, and the origin is probably the South Africa. Nowadays, this species can be found in a latitudinal range of 35º, from Uruguay to Belem (Brazil). Esterases comprise a multi-functional and heterogeneous group of enzymes that participated in ester hydrolysis. In insects, they are related to several metabolic processes, including the reproductive function. Esterase patterns in Z. indianus and Drosophila melanogaster were characterized in polyacrylamide gels (PAGE). Two - esterases from Z. indianus, EST-2 and EST-5, showed similarity preference for substrate - naftil acetate and patterns of inhibition as compared to the EST-6 of D. melanogaster, suggesting a possible role in reproductive biology for both enzymes. Biochemical characterization of these esterases and its differential expression in males, suggest orthology among their genes. In this work, the genomic DNA from Z. indianus was submitted to PCR experiments using 3 sets of D. melanogaster Est-6 sequence primers. The PCR products were directly sequenced; its GC content was 48.5% and presented a 73% similarity compared to Est-6 D. melanogaster sequence. Analysis of sequence data, by estimates of nonsynonymous/synonymous rate ratios, showed that the majority of sites evolves under strong or moderate negative selection (81%) and a minority of sites (19%). is under significant positive selection. Molecular modeling indicates that the fundamental structural features important for catalysis are conserved in the esterase of Z. indianus and human bilesalt activated lipase (BAL), which was used as structural model. Structural and sequence comparisons suggest the evolutionary relationship between ...(Complete abstract click electronic access below)
Descrição
Palavras-chave
Drosófila - Genética, Esterases, Drosophila melanogaster, Zaprionus indianus, Male esterase, Esterase-6
Como citar
PAULINO, Rafael Marques. Caracterização molecular e bioquímica de uma esterase macho-específica em Zaprionus indianus. 2008. 77 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Biociências, Letras e Ciências Exatas, 2008.