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利用RNA测序分析确定里氏木霉中CRE1在碳代谢物阻遏过程中的全基因组作用。

Defining the genome-wide role of CRE1 during carbon catabolite repression in Trichoderma reesei using RNA-Seq analysis.

作者信息

Antoniêto Amanda Cristina Campos, dos Santos Castro Lílian, Silva-Rocha Rafael, Persinoti Gabriela Felix, Silva Roberto Nascimento

机构信息

Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.

Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, São Paulo, Brazil.

出版信息

Fungal Genet Biol. 2014 Dec;73:93-103. doi: 10.1016/j.fgb.2014.10.009. Epub 2014 Oct 18.

Abstract

The ascomycete Trichoderma reesei is one of the most well-studied cellulolytic fungi and is widely used by the biotechnology industry in the production of second generation bioethanol. The carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1. CCR represses genes related to cellulase production when a carbon source is readily available in the medium. Using RNA sequencing, we investigated CCR during the synthesis of cellulases, comparing the T. reesei Δcre1 mutant strain with its parental strain, QM9414. Of 9129 genes in the T. reesei genome, 268 genes were upregulated and 85 were downregulated in the presence of cellulose (Avicel). In addition, 251 genes were upregulated and 230 were downregulated in the presence of a high concentration of glucose. Genes encoding cellulolytic enzymes and transcription factors and genes related to the transport of nutrients and oxidative metabolism were also targets of CCR, mediated by CRE1 in a carbon source-dependent manner. Our results also suggested that CRE1 regulates the expression of genes related to the use of copper and iron as final electron acceptors or as cofactors of enzymes that participate in biomass degradation. As a result, the final effect of CRE1-mediated transcriptional regulation is to modulate the access of cellulolytic enzymes to cellulose polymers or blocks the entry of cellulase inducers into the cell, depending on the glucose content in the medium. These results will contribute to a better understanding of the mechanism of carbon catabolite repression in T. reesei, thereby enhancing its application in several biotechnology fields.

摘要

子囊菌里氏木霉是研究最为深入的纤维素分解真菌之一,被生物技术产业广泛用于生产第二代生物乙醇。里氏木霉采用的碳分解代谢物阻遏(CCR)机制由转录因子CRE1介导。当培养基中存在易于利用的碳源时,CCR会抑制与纤维素酶产生相关的基因。我们利用RNA测序技术,在纤维素酶合成过程中研究了CCR,将里氏木霉Δcre1突变菌株与其亲本菌株QM9414进行了比较。在里氏木霉基因组的9129个基因中,有268个基因在纤维素(微晶纤维素)存在时上调,85个基因下调。此外,在高浓度葡萄糖存在时,有251个基因上调,230个基因下调。编码纤维素分解酶和转录因子的基因以及与营养物质转运和氧化代谢相关的基因也是CCR的作用靶点,由CRE1以碳源依赖的方式介导。我们的结果还表明,CRE1调节与将铜和铁用作最终电子受体或参与生物质降解的酶的辅因子相关的基因表达。因此,CRE1介导的转录调控的最终作用是根据培养基中的葡萄糖含量,调节纤维素分解酶与纤维素聚合物的接触,或阻止纤维素酶诱导剂进入细胞。这些结果将有助于更好地理解里氏木霉中碳分解代谢物阻遏的机制,从而加强其在多个生物技术领域的应用。

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