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Cellulase gene expression in the model cellulolytic fungus Trichoderma reesei is supposed to be controlled by an intricate regulatory network involving multiple transcription factors. Here, we identified a novel transcriptional repressor of cellulase gene expression, Rce1. Disruption of the rce1 gene not only facilitated the induced expression of cellulase genes but also led to a significant delay in terminating the induction process. However, Rce1 did not participate in Cre1-mediated catabolite repression. Electrophoretic mobility shift (EMSA) and DNase I footprinting assays in combination with chromatin immunoprecipitation (ChIP) demonstrated that Rce1 could bind directly to a cbh1 (cellobiohydrolase 1-encoding) gene promoter region containing a cluster of Xyr1 binding sites. Furthermore, competitive binding assays revealed that Rce1 antagonized Xyr1 from binding to the cbh1 promoter. These results indicate that intricate interactions exist between a variety of transcription factors to ensure tight and energy-efficient regulation of cellulase gene expression in T. reesei. This study also provides important clues regarding increased cellulase production in T. reesei.
Mol Microbiol 2017 07
PMID:Rce1, a novel transcriptional repressor, regulates cellulase gene expression by antagonizing the transactivator Xyr1 in Trichoderma reesei. 2837 98

Testing of cellulases on real biomass samples is required to do a true assessment of their efficacy for biomass degradation. Cellulase enzymes belong to a number of different glycosyl hydrolase families, all with different activity, specificity and modes of action. The concerted and synergistic action of these different cellulases determines the efficacy for plant cell wall deconstruction and cellulose hydrolysis. However, the plant cell wall of lignocellulosic materials is a very complex matrix and the efficacy of a cellulase preparation to degrade lignocellulosic materials is influenced by many factors. In this chapter, two protocols for testing efficacy of cellulases on pretreated biomass samples are described. The first protocol describes a small-scale setup employing low solids concentration that easily enables the testing of a larger number of samples. The second protocol describes a method for testing the efficacy of cellulases at conditions more closely resembling industrial conditions, i.e., high solids concentrations. Both protocols can be used to test the cellulases under a variety of substrate types, substrate concentrations, enzyme loadings and process conditions. The protocols can also be used to evaluate different feedstocks.
Methods Mol Biol 2018
PMID:Test of Efficacy of Cellulases for Biomass Degradation. 2985 61

Cellulase gene expression in Trichoderma reesei is highly responsive to environmental cues and is under stringent regulation by multiple transcription factors. XYR1 (Xylanase regulator 1) has been identified as the most important transcriptional activator of cellulase/hemicellulase gene expression although the precise transactivating mechanism remains largely elusive. Here we show that the activation domain of XYR1 interacts with the T. reesei homolog of the TrSNF12 subunit of SWI/SNF complex. Deletion of Trsnf12 markedly impaired the induced cellulase gene expression. Individual loss of other SWI/SNF subunits including the catalytic subunit also severely compromised cellulase gene expression and interfered with loss of histone H4 in the cbh1 and eg1 promoters upon cellulose induction. In addition, we find that the SWI/SNF occupancy on cellulase gene promoters strictly required XYR1 and TrSNF12 but TrSNF12 was dispensable for the XYR1 binding to these promoters. These data suggest a model in which XYR1 recruits SWI/SNF through direct interactions with TrSNF12 to remodel chromatin at cellulase gene promoters, thereby activating cellulase gene expression to initiate the cellulolytic response in T. reesei.
Mol Microbiol 2019 10
PMID:Trichoderma reesei XYR1 recruits SWI/SNF to facilitate cellulase gene expression. 3130 4


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