School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, No. 382, Waihuan East Rd, Guangzhou, 510006, China.
Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, 510006, China.
Biotechnol Lett. 2020 Apr;42(4):613-624. doi: 10.1007/s10529-020-02806-3. Epub 2020 Jan 22.
To survey genome-scale protease profiles regulated by the Aspergillus niger transcription factor PrtT and further controlled by carbon sources.
The PrtT disruption mutant (delprtT) and overexpression (OEprtT) strains were successfully generated and further confirmed by phenotypic and protease activity analysis. RNA-seq analysis of WT and mutants identified 32 differentially expressed protease genes, which mostly belonged to serine-type peptidases, aspartic-type endopeptidases, aminopeptidases and carboxypeptidases. Furthermore, based on the MEME predicted motif analysis of the PrtT promoter, EMSA and phenotypic and qRT-PCR analyses confirmed that the carbon metabolism regulator AmyR directly regulated the protease genes and their regulatory factor PrtT.
Thirty-two PrtT-regulated protease genes were identified by RNA-seq, and the secondary carbon source regulator AmyR was found to have a negative regulatory effect on the expression of PrtT and its target protease genes.
调查黑曲霉转录因子 PrtT 调控的基因组规模蛋白酶谱，并进一步受碳源控制。
成功构建了 PrtT 缺失突变体（delprtT）和过表达（OEprtT）菌株，并通过表型和蛋白酶活性分析进一步证实。WT 和突变体的 RNA-seq 分析鉴定了 32 个差异表达的蛋白酶基因，这些基因主要属于丝氨酸型肽酶、天冬氨酸内肽酶、氨肽酶和羧肽酶。此外，基于 PrtT 启动子的 MEME 预测基序分析、EMSA 以及表型和 qRT-PCR 分析证实，碳代谢调节剂 AmyR 直接调控蛋白酶基因及其调控因子 PrtT。
通过 RNA-seq 鉴定了 32 个受 PrtT 调控的蛋白酶基因，发现次级碳源调节剂 AmyR 对 PrtT 及其靶标蛋白酶基因的表达具有负调控作用。
