Regulation of cellulase enzyme expression in Trichoderma

该项目旨在为丝状真菌里氏木霉纤维素酶的转录调控提供新的信息。T.里氏木霉是一种被广泛商业开发用于生产天然（主要是纤维素酶）酶和越来越多的异源蛋白质的真菌。对T. reesei可查阅（http：//genome.jgi-psf.org/Trire2/Trire2.home.html）。虽然T. Reesei基因组编码几种纤维素酶的表达，CBH 1基因在其表达方面已经被最广泛地研究，并且CBH 1启动子用于驱动异源蛋白的表达。该项目将建立在我们未发表的数据基础上，这些数据首次显示了染色质在其调控中的重要性，包括诱导和抑制过程中核小体定位的变化。此外，我们有初步的数据显示蛋白质结合的ORF在诱导过程中，表明ORF参与调控除了启动子区域。这在丝状真菌中是前所未有的。该项目的总体目标是确定丝状真菌中靶基因转录控制的新调控原则。该项目的具体实验目的是确认在诱导和抑制过程中chb 1启动子和ORF的核小体重排，并指定已知调节因子（例如Cre 1，Ace 1）在核小体重排中的作用。第二个目的是定义一个模型的调节cbh 1。该项目还将探索蛋白质结合ORF在转录调控过程中的作用。可用于探索核小体定位的方法（包括分离染色质、核酸酶敏感性测定和杂交研究）、诱导和阻遏的培养条件（和中性条件）以及用于探索某些转录因子作用的某些突变株，都在诺丁汉或Roal研究机构中进行。该项目为确定真菌中转录调控的新机制提供了令人兴奋的可能性。学生将接受微生物学，基因组学和分子生物学的培训，并将有机会与诺丁汉的数学家进行交流，以确定cbh 1基因调控的模型。该模型可以进行测试，并使用基因阵列方法对真菌整体的基因调控进行相关性检查，该方法最近才用于T。reesei。这一新的信息对Roal公司来说很重要，Roal公司是一家利用丝状真菌的公司，包括T。瑞思，在其业务。

The project aims to provide new information on the transcriptional regulation of expression of cellulase in the filamentous fungus Trichoderma reesei. T. reesei is a fungus exploited extensively commercially for production of native (mainly cellulases) enzymes and, increasingly, heterologous proteins. The genome sequence of T. reesei is available (http://genome.jgi-psf.org/Trire2/Trire2.home.html). Although the T. reesei genome encodes for the expression of several cellulases, the cbh1 gene has been the most extensively studied in relation to its expression and the cbh1 promoter is used to drive expression of heterologous proteins. This project will build upon our unpublished data which show, for the first time, the importance of chromatin in its regulation, including changes in nucleosomal positioning during induction and repression. Moreover, we have preliminary data showing protein binding to the ORF during induction, indicating that the ORF is involved in regulation in addition to the promoter region. That is unprecedented in filamentous fungi. The overall Aim of the project is to define new regulatory principles in the transcriptional control of a target gene in a filamentous fungus. The specific experimental Aims of the project are to confirm the nucleosomal rearrangements of the chb1 promoter and ORF during induction and repression and to specify the roles, if any, of known regulators (e.g. Cre1, Ace1) in the nucleosomal rearrangements. A secondary Aim is to define a model for the regulation of cbh1. The project will also explore the role of protein binding to the ORF during transcriptional regulation. The methodologies available for exploration of nucleosomal positioning (involving the isolation of chromatin, nuclease sensitivity assays, and hybridisation studies), culture conditions for induction and repression (and neutral conditions), and some mutant strains for exploration of the roles of some transcription factors are all in place in Nottingham or the research facility of Roal. This project provides an exciting possibility for defining new mechanisms of transcriptional regulation in fungi. The student will receive training in microbiology, genomics and molecular biology and will have the opportunity to interface with mathematicians in Nottingham in order to define a model for the regulation of the cbh1 gene That model can be tested and its relevance to gene regulation in the fungus as a whole examined using gene array methodology, which is only recently available for T. reesei. This new information will be important for Roal, a company that exploits filamentous fungi, including T. reesei, in its business.