Identification of Aspergillus brlA response elements (BREs) by genetic selection in yeast.
Identification of Aspergillus brlA response elements (BREs) by genetic selection in yeast.
复制标题
通过酵母遗传选择鉴定曲霉 brlA 响应元件 (BRE)。
DOI：
10.1093/genetics/133.1.29
复制
发表时间：
1993
期刊：
影响因子：
3.3
通讯作者：
Timberlake,WE
中科院分区：
文献类型：
--
作者：
Chang,YC;Timberlake,WE
构巢曲霉的 brlA 基因在控制分生孢子梗发育中发挥着核心作用。为了测试 brlA 编码转录调节因子的假设并鉴定 BrlA 多肽的相互作用位点，我们在含有插入到与大肠杆菌 lacZ 基因融合的最小酵母启动子上游的曲霉属 DNA 序列的酿酒酵母（酵母）菌株中表达 brlA。最初，对来自发育调节的 rodA 基因启动子区域的 DNA 片段进行了测试，结果显示其介导 brlA 依赖性转录激活。根据曲霉属基因组文库对酵母中 brlA 的反应能力，从曲霉属基因组文库中选择了另外两个 DNA 片段。这些片段包含存在于 rodA 片段中的序列基序的多个拷贝，我们建议将其作为 BrlA 相互作用的位点并指定为 brlA 响应元件 (BRE)。含有位于最小曲霉属启动子上游的 BRE 的 DNA 片段能够赋予曲霉属发育调节作用。 rodA 上游区域 BRE 的缺失大大降低了其发育诱导。合成寡核苷酸的多个拷贝，具有在酵母中 BRE 介导的 brlA 依赖性转录激活中鉴定的共有序列。结果表明，brlA 的主要活性是转录激活，并初步鉴定了 BrlA 多肽的相互作用位点。
The brlA gene of Aspergillus nidulans plays a central role in controlling conidiophore development. To test the hypothesis that brlA encodes a transcriptional regulator and to identify sites of interaction for the BrlA polypeptide, we expressed brlA in Saccharomyces cerevisiae (yeast) strains containing Aspergillus DNA sequences inserted upstream of a minimal yeast promoter fused to the Escherichia coli lacZ gene. Initially, a DNA fragment from the promoter region of the developmentally regulated rodA gene was tested and shown to mediate brlA-dependent transcriptional activation. Two additional DNA fragments were selected from an Aspergillus genomic library by their ability to respond to brlA in yeast. These fragments contained multiple copies of a sequence motif present in the rodA fragment, which we propose to be sites for BrlA interaction and designate brlA response elements (BREs). DNA fragments containing BREs upstream of a minimal Aspergillus promoter were capable of conferring developmental regulation in Aspergillus. Deletion of BREs from the upstream region of rodA greatly decreased its developmental induction. Multiple copies of a synthetic oligonucleotide with the consensus sequence identified among the BREs mediated brlA-dependent transcriptional activation in yeast. The results show that a primary activity of brlA is transcriptional activation and tentatively identify sites of interaction for the BrlA polypeptide.