Metabolismo de glicogênio e relógio biológico em Neurospora crassa: fatores e cofatores de transcrição envolvidos nos processos
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Data
2012-08-15
Autores
Virgilio, Stela [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O fungo filamentoso Neurospora crassa é um organismo modelo utilizado na compreensão de diversos aspectos da biologia dos eucariotos, e tem sido usado, em nosso laboratório, para estudos celulares básicos, como os mecanismos bioquímicos e moleculares envolvidos na regulação do metabolismo de glicogênio. Uma análise sistemática realizada com uma coleção de linhagens mutantes em genes codificadores de fatores de transcrição permitiu identificar várias proteínas potencialmente envolvidas na regulação do metabolismo do glicogênio neste organismo. Algumas linhagens mutantes apresentaram alterações no perfil de acúmulo de glicogênio e na expressão dos genes que codificam as enzimas glicogênio sintase (gsn) e glicogênio fosforilase (gpn) quando comparadas à linhagem selvagem. Dentre estas, duas linhagens mutantes em genes que codificam para a proteína RCO-1 (regulator of conidiation-1) e para uma proteína hipotética foram selecionadas para o presente estudo, levando em consideração que ambas linhagens também apresentaram variações na progressão do ciclo celular quando analisadas por citometria de fluxo. Como a proteína RCO-1 é uma provável parceira da proteína RCM-1 (regulator of conidiation and morphology-1), então a linhagem mutante no gene codificador de RCM-1 foi incluída neste trabalho. Portanto, foi feita a caracterização de um fator de transcrição anotado como proteína hipotética e de dois cofatores transcricionais RCO-1 e RCM-1, ortólogos ao complexo corepressor Tup1-Ssn6 de Saccharomyces cerevisiae. As proteínas RCO-1, RCM-1 e a codificada pela ORF NCU09739 estão envolvidas na regulação do metabolismo do glicogênio, atuando na regulação da expressão dos genes gsn e/ou gpn. Estas mesmas proteínas também são necessárias para o crescimento e desenvolvimento normal do...
The filamentous fungus Neurospora crassa is a model organism used to understand various aspects of eukaryotic biology. It has been used, in our laboratory, in basic cellular studies, such as the biochemical and molecular mechanisms involved in the regulation of glycogen metabolism. A systematic analysis performed with a collection of mutant strains in genes encoding transcription factors led to the identification of proteins likely involved in the regulation of glycogen metabolism in this organism. Some mutant strains showed changes in the glycogen accumulation profile and in the expression of the genes encoding the enzymes glycogen synthase (gsn) and glycogen phosphorylase (gpn) when compared to the wild-type strain. Among these, two mutant strains in the genes encoding RCO-1 (regulator of conidiation-1) and a hypothetical proteins were selected for the present study. Both strains presented variations in cell cycle progression when analyzed by flow cytometry. RCO-1 protein is likely a partner of RCM-1 (regulator of conidiation and morphology-1) protein, thus the mutant strain in the gene encoding RCM-1 was included in this work. Therefore, we performed the characterization of a transcription factor annotated as a hypothetical protein and the two transcriptional cofactors RCO-1 and RCM-1, orthologs of the Saccharomyces cerevisiae corepressor complex Tup1-Ssn6. RCO-1, RCM-1 and the product of the ORF NCU09739 are involved in the regulation of glycogen metabolism, acting in the regulation of gsn and/or gpn gene expression. The same proteins are necessary for growth and normal development of the fungus, since the mutant strains showed changes in hyphae length, pigmentation and conidiation. Gene expression analysis showed that the NCU09739 gene was highly expressed at the beginning of the conidia germination, showing the importance... (Complete abstract click electronic access below)
The filamentous fungus Neurospora crassa is a model organism used to understand various aspects of eukaryotic biology. It has been used, in our laboratory, in basic cellular studies, such as the biochemical and molecular mechanisms involved in the regulation of glycogen metabolism. A systematic analysis performed with a collection of mutant strains in genes encoding transcription factors led to the identification of proteins likely involved in the regulation of glycogen metabolism in this organism. Some mutant strains showed changes in the glycogen accumulation profile and in the expression of the genes encoding the enzymes glycogen synthase (gsn) and glycogen phosphorylase (gpn) when compared to the wild-type strain. Among these, two mutant strains in the genes encoding RCO-1 (regulator of conidiation-1) and a hypothetical proteins were selected for the present study. Both strains presented variations in cell cycle progression when analyzed by flow cytometry. RCO-1 protein is likely a partner of RCM-1 (regulator of conidiation and morphology-1) protein, thus the mutant strain in the gene encoding RCM-1 was included in this work. Therefore, we performed the characterization of a transcription factor annotated as a hypothetical protein and the two transcriptional cofactors RCO-1 and RCM-1, orthologs of the Saccharomyces cerevisiae corepressor complex Tup1-Ssn6. RCO-1, RCM-1 and the product of the ORF NCU09739 are involved in the regulation of glycogen metabolism, acting in the regulation of gsn and/or gpn gene expression. The same proteins are necessary for growth and normal development of the fungus, since the mutant strains showed changes in hyphae length, pigmentation and conidiation. Gene expression analysis showed that the NCU09739 gene was highly expressed at the beginning of the conidia germination, showing the importance... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Biotecnologia, Bioquímica, Glicogenio, Expressão gênica, Proteínas, Glycogen
Como citar
VIRGILIO, Stela. Metabolismo de glicogênio e relógio biológico em Neurospora crassa: fatores e cofatores de transcrição envolvidos nos processos. 2012. 138 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Química, 2012.