Mello-de-Sousa Thiago M, Rassinger Alice, Derntl Christian, Poças-Fonseca Marcio J, Mach Robert L, Mach-Aigner Astrid R
Department for Biotechnology and Microbiology, TU Wien, Vienna, Austria;
Department of Genetics and Morphology, Institute of Biological Sciences, University of Brasilia, Brasília-DF, Brazil.
Curr Genomics. 2016 Apr;17(2):145-52. doi: 10.2174/1389202917666151116211812.
The ascomycete Trichoderma reesei is used for the production of plant cell wall-degrading enzymes in industrial scale. The interplay of the transactivator Xyr1 and the repressor Cre1 mainly regulates the expression of these enzymes. During induc-ing conditions, such as in the presence of sophorose, the transcription of the two major cellulase-encoding genes, cbh1 and cbh2, is activated as well as the expression of xyr1. In the presence of D-glucose carbon catabolite repression mediated by Cre1 takes place and the expression of Xyr1 and the plant cell wall-degrading enzymes is down-regulated. In this study we compare the chromatin status of xyr1, cbh1, and cbh2 promoters in the wild-type strain and the Cre1-deficient strain Rut-C30. Chromatin rearrangement occurs in the xyr1 promoter during induction on sophorose. Chromatin opening and protein-DNA interactions in the xyr1 promoter were detected especially in a region located 0.9 kb upstream the translation start co-don, which bears several putative Cre1-binding sites and a CCAAT-box. Moreover, the xyr1 promoter is overall more acces-sible in a cre1-truncated background, no matter which carbon source is present. This makes the xyr1 regulatory sequence a good target for promoter engineering aiming at the enhancement of cellulase production.
子囊菌里氏木霉用于大规模生产植物细胞壁降解酶。反式激活因子Xyr1和阻遏蛋白Cre1的相互作用主要调节这些酶的表达。在诱导条件下,例如在槐糖存在的情况下,两个主要纤维素酶编码基因cbh1和cbh2的转录以及xyr1的表达都会被激活。在D-葡萄糖存在的情况下,由Cre1介导的碳代谢物阻遏发生,Xyr1和植物细胞壁降解酶的表达被下调。在本研究中,我们比较了野生型菌株和Cre1缺陷型菌株Rut-C30中xyr1、cbh1和cbh2启动子的染色质状态。在槐糖诱导过程中,xyr1启动子发生染色质重排。特别是在翻译起始密码子上游0.9 kb的区域检测到xyr1启动子的染色质开放和蛋白质-DNA相互作用,该区域有几个假定的Cre1结合位点和一个CCAAT框。此外,无论存在何种碳源,xyr1启动子在cre1截短的背景下总体上更容易接近。这使得xyr1调控序列成为旨在提高纤维素酶产量的启动子工程的良好靶点。