The transcription factor PrtT and its target protease profiles in Aspergillus niger are negatively regulated by carbon sources
The transcription factor PrtT and its target protease profiles in Aspergillus niger are negatively regulated by carbon sources
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黑曲霉中的转录因子 PrtT 及其靶蛋白酶谱受到碳源的负调控
DOI：
10.1007/s10529-020-02806-3
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发表时间：
2020-01-22
影响因子：
2.7
通讯作者：
Pan, Li
中科院分区：
文献类型：
--
作者：
Huang, Lianggang;Dong, Liangbo;Pan, Li
目的研究黑曲霉转录因子PrtT调控并进一步受碳源调控的蛋白酶基因组图谱。结果成功生成了PrtT断裂突变株（delprtT）和过表达株（OEprtT），并通过表型和蛋白酶活性分析进一步证实了这两株菌株的存在。WT和突变体的RNA-seq分析鉴定出32个差异表达的蛋白酶基因，主要属于丝氨酸型肽酶、天冬氨酸型内肽酶、氨基肽酶和羧基肽酶。此外，基于PrtT启动子的MEME预测基序分析、EMSA和表型分析以及qRT-PCR分析证实，碳代谢调节因子AmyR直接调控蛋白酶基因及其调控因子PrtT。结论通过RNA-seq鉴定出32个PrtT调控的蛋白酶基因，发现次级碳源调控因子AmyR对PrtT及其靶蛋白酶基因的表达具有负向调控作用。
Objective To survey genome-scale protease profiles regulated by the Aspergillus niger transcription factor PrtT and further controlled by carbon sources. Results 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. Conclusion 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.