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米曲霉分泌应激下抑制米曲霉淀粉酶A基因所需功能性顺式元件的鉴定。

Identification of functional cis-elements required for repression of the Taka-amylase A gene under secretion stress in Aspergillus oryzae.

作者信息

Zhou Bin, Wang Chao, Wang Bin, Li Xiupeng, Xiao Jing, Pan Li

机构信息

School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, People's Republic of China.

出版信息

Biotechnol Lett. 2015 Feb;37(2):333-41. doi: 10.1007/s10529-014-1691-2. Epub 2014 Oct 4.

Abstract

The expression of secreted proteins in filamentous fungi is down-regulated by a transcriptional feedback mechanism under endoplasmic reticulum stress, termed repression under secretion stress (RESS). To investigate the RESS mechanism, we analyzed the expression of the Taka-amylase A gene (amyB) in Aspergillus oryzae, which was depressed under secreted protein stress. We conducted a truncation and deletion analysis of the amyB promoter to identify cis-elements required for RESS. A nucleotide sequence (positions -378 to -291) without any binding sites for the transcriptional activator AmyR, which is involved in amylolytic gene expression, was required for RESS. The octamer sequence TCACGGGC (positions -307 to -300) constituted the core sequence of the upstream activating element essential for amyB down-regulation under secretion stress. Both the inactivation of AmyR and RESS contributed to the down-regulation of amyB expression under ER stress.

摘要

丝状真菌中分泌蛋白的表达在内质网应激下通过一种转录反馈机制被下调,这种机制被称为分泌应激下的抑制(RESS)。为了研究RESS机制,我们分析了米曲霉中Taka淀粉酶A基因(amyB)的表达,该基因在分泌蛋白应激下会受到抑制。我们对amyB启动子进行了截短和缺失分析,以确定RESS所需的顺式元件。RESS需要一个核苷酸序列(位置-378至-291),该序列没有参与淀粉分解基因表达的转录激活因子AmyR的任何结合位点。八聚体序列TCACGGGC(位置-307至-300)构成了分泌应激下amyB下调所必需的上游激活元件的核心序列。AmyR的失活和RESS都导致了内质网应激下amyB表达的下调。

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