MtTRC-1, a Novel Transcription Factor, Regulates Cellulase Production via Directly Modulating the Genes Expression of the Mthac-1 and Mtcbh-1 in Myceliophthora thermophila.
MtTRC-1, a Novel Transcription Factor, Regulates Cellulase Production via Directly Modulating the Genes Expression of the Mthac-1 and Mtcbh-1 in Myceliophthora thermophila.
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DOI：
10.1128/aem.01263-22
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发表时间：
2022-10-11
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
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嗜热真菌Myceliophthora thermophila已被用于生产工业酶和生物基化学品。在腐食性真菌中，纤维素酶的调控机制已被研究，揭示了多个转录因子的参与。然而，在M。在嗜热菌中，影响纤维素酶基因表达和分泌的转录因子仍然很大程度上未知。在这项研究中，我们确定并表征了一个新的纤维素酶调节剂（MtTRC-1）在M。通过功能基因组学和遗传分析相结合的嗜热菌。Mttrc-1的缺失导致纤维素酶的产量和活性显著降低。转录组分析显示，在纤维素酶诱导条件下，MtTRC-1的破坏不仅导致了主要纤维素酶编码基因的下调，还导致了UPR途径转录调控因子MtHAC-1的下调。在此，我们还表征了M中酵母HAC 1 p的直系同源物。嗜热菌我们发现，Mthac-1 mRNA经历了内质网（ER）应力诱导的剪接，通过删除一个23个核苷酸（nt）的内含子。值得注意的是，纤维素上的蛋白分泌由于MtHAC-1的缺失而显著受损。此外，在不存在MtHAC-1的情况下，在各种碳源上的菌落生长是有缺陷的。电泳迁移率变动分析和染色质免疫沉淀分析证实MtTRC-1在体外和体内通过直接与启动子结合来调节Mthac-1和主要纤维素酶基因Mtcbh-1的转录。此外，DNase I足迹分析鉴定了推定的共有结合位点（5′-GNG/C-3′）。这些结果揭示了MtTRC-1对于正调节纤维素酶生产的重要性。这一发现阐明了纤维素分解酶生产中涉及的复杂调控途径。在本研究中，我们在M.嗜热菌，其通过直接转录调节Mthac-1和Mtcbh-1基因来调节纤维素酶的产生。结果表明，MtHAC-1是影响M.嗜热菌研究结果表明，该嗜热真菌通过一个多层次网络调控纤维素酶的合成，其中蛋白质分泌途径受MtHAC-1依赖的UPR途径调控，纤维素酶基因表达受转录因子直接调控。Mttrc 1在丝状真菌中的保守性表明，这种机制可能被利用来工程化能够在工业规模上生产蛋白质的丝状真菌细胞工厂。
The thermophilic fungus Myceliophthora thermophila has been used to produce industrial enzymes and biobased chemicals. In saprotrophic fungi, the mechanisms regulating cellulase production have been studied, which revealed the involvement of multiple transcription factors. However, in M. thermophila, the transcription factors influencing cellulase gene expression and secretion remain largely unknown. In this study, we identified and characterized a novel cellulase regulator (MtTRC-1) in M. thermophila through a combination of functional genomics and genetic analyses. Deletion of Mttrc-1 resulted in significantly decreased cellulase production and activities. Transcriptome analysis revealed downregulation of not only the encoding genes of main cellulases but also the transcriptional regulator MtHAC-1 of UPR pathway after disruption of MtTRC-1 under cellulolytic induction conditions. Herein, we also characterized the ortholog of the yeast HAC1p in M. thermophila. We show that Mthac-1 mRNA undergoes an endoplasmic reticulum (ER) stress-induced splicing by removing a 23-nucleotide (nt) intron. Notably, the protein secretion on cellulose was dramatically impaired by the deletion of MtHAC-1. Moreover, the colonial growth on various carbon sources was defective in the absence of MtHAC-1. Electrophoretic mobility shift assays and chromatin immunoprecipitation assays verified MtTRC-1 regulates the transcription of Mthac-1 and the major cellulase gene Mtcbh-1 by binding directly to the promoters in vitro and in vivo. Furthermore, DNase I footprinting assays identified the putative consensus binding site (5′-GNG/C-3′). These results revealed the importance of MtTRC-1 for positively regulating cellulase production. This finding has clarified the complex regulatory pathways involved in cellulolytic enzyme production. IMPORTANCE In the present study, we characterized a novel regulator MtTRC-1 in M. thermophila, which regulated cellulase production through direct transcriptional regulation of the Mthac-1 and Mtcbh-1 genes. Our data demonstrated that MtHAC-1 is a key factor for the cellulase secretion capacity of M. thermophila. Our data indicate that this thermophilic fungus regulates cellulase production through a multilevels network, in which the protein secretory pathway is modulated by MtHAC-1-dependent UPR pathway and the cellulase gene expression is directly regulated in parallel by transcription factors. The conservation of Mttrc1 in filamentous fungi suggests this mechanism may be exploited to engineer filamentous fungal cell factories capable of producing proteins on an industrial scale.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
6.4
作者：
Craig JP;Coradetti ST;Starr TL;Glass NL
通讯作者：
Glass NL
影响因子：
3.6
作者：
Benz JP;Chau BH;Zheng D;Bauer S;Glass NL;Somerville CR
通讯作者：
Somerville CR
影响因子：
3.6
作者：
Al-Sheikh, H;Watson, AJ;Archer, DB
通讯作者：
Archer, DB
里氏木霉 Rut-C30 中 Ca2 响应信号通路调节纤维素酶表达和分泌的表征
影响因子：
3.6
作者：
Chen, Ling;Zou, Gen;Zhou, Zhihua
通讯作者：
Zhou, Zhihua