4.4 Article

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

期刊

BIOTECHNOLOGY LETTERS
卷 37, 期 2, 页码 333-341

出版社

SPRINGER
DOI: 10.1007/s10529-014-1691-2

关键词

Aspergillus oryzae; Quantitative real-time PCR; Repression under secretion stress; Taka-amylase A gene promoter; Truncation and deletion analysis

资金

  1. National Science and Technology Foundation in China [2011AA100905]
  2. Guangdong Provincial Department of Science and Technology Research Project [2012 B010900028]
  3. National Natural Science Foundation of China [31100026]

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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.

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