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CLP1 a Novel Plant Homeo Domain Protein Participates in Regulating Cellulase Gene Expression in the Filamentous Fungus Trichoderma reesei

机译:CLP1一种新型植物同源域蛋白参与调节丝状真菌里氏木霉中的纤维素酶基因表达。

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摘要

The stringent regulatory network of cellulase gene expression in the filamentous fungus Trichoderma reesei involves multiple transcriptional regulators. However, identification and mechanistic investigation of these regulators are still insufficient. Here, we identified a novel transcriptional regulator, CLP1, a plant homeo domain (PHD) Protein that participates in regulating T. reesei cellulase gene expression. Phylogenetic analyses demonstrated that CLP1 homologs are widely distributed in filamentous fungi including Trichoderma, Penicillium, Fusarium, Neurospora, and Aspergillus species. We demonstrated that CLP1 is a nuclear protein and lack of CLP1 significantly impaired the induced expression of cellulase genes. ChIP experiments showed CLP1 binding to the cellulase gene promoters specifically under cellulose conditions and compromised XYR1 occupancy on the same promoters in the absence of CLP1 at the early induction stage. XYR1 overexpression fully rescued the defect in cellulase production but not the defect in conidia formation in the clp1 null mutant. Further analysis showed that the PHD is required for the CLP1 appropriate subcellular localization as well as the induced cellulase gene expression and conidiation. Taken together, these data demonstrated an important role of CLP1 in the regulation of cellulase and xylanase gene expression in T. reesei.
机译:丝状真菌里氏木霉中纤维素酶基因表达的严格调控网络涉及多个转录调控因子。但是,对这些调节器的识别和机理研究仍然不够。在这里,我们确定了一种新型的转录调节因子CLP1,一种植物同源域(PHD)蛋白,参与调节里氏木霉纤维素酶基因的表达。系统发育分析表明,CLP1同源物广泛分布于丝状真菌中,包括木霉菌,青霉菌,镰刀菌,神经孢菌和曲霉菌。我们证明了CLP1是一种核蛋白,缺少CLP1会大大削弱纤维素酶基因的诱导表达。 ChIP实验显示,CLP1在纤维素条件下特别是与纤维素酶基因启动子结合,并且在诱导初期,在没有CLP1的情况下,相同启动子上的XYR1占有率受损。 XYR1过表达完全挽救了纤维素酶生产中的缺陷,但不能挽救clp1 null突变体中分生孢子形成中的缺陷。进一步的分析表明,PHD是CLP1合适的亚细胞定位以及诱导的纤维素酶基因表达和分支的必需条件。综上所述,这些数据证明了CLP1在里氏木霉中纤维素酶和木聚糖酶基因表达的调节中的重要作用。

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