<br>Yas3p 是一种 Opi1 家族转录因子，可响应解脂耶氏酵母中的正构烷烃调节细胞色素 P450 表达。,Journal of Biological Chemistry

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Yas3p 是一种 Opi1 家族转录因子，可响应解脂耶氏酵母中的正构烷烃调节细胞色素 P450 表达。
Journal of Biological Chemistry ( IF 4.0 ) Pub Date : 2009 Mar 13 , DOI: 10.1074/jbc.m806864200
Kiyoshi Hirakawa ₁ , Satoshi Kobayashi , Takuro Inoue , Setsu Endoh-Yamagami , Ryouichi Fukuda , Akinori Ohta

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在烷烃同化酵母解脂耶氏酵母中，ALK1 是一种编码细胞色素 P450 的基因，可催化正烷烃氧化的第一步，其表达是由正烷烃诱导的。我们之前证明，两种基本的螺旋-环-螺旋蛋白 Yas1p 和 Yas2p 通过形成异质复合物并与烷烃响应元件 1 (ARE1) 结合，以烷烃依赖性方式激活 ALK1 的转录，ARE1 是一种顺式作用元件。 ALK1 启动子。在这里，我们鉴定了 Opi1 家族转录因子 Yas3p，参与 ALK 基因的烷烃依赖性转录调控。在葡萄糖、甘油和正烷烃上生长的细胞中，YAS3 的缺失导致 ALK1 mRNA 显着增加。 YAS3 缺失还导致甘油和正癸烷上含有 ARE1 的启动子驱动的报告基因表达显着升高。在体外，细菌表达的 Yas3p 与 Yas2p 特异性结合，但不与 Yas1p 结合。此外，虽然绿色荧光蛋白标记的Yas3p在含葡萄糖的培养基中定位于细胞核中，但在转移到含有正癸烷的培养基中后，其定位改变为内质网样区室。这些发现表明 Yas3p 作为转录反应的主要调节因子发挥作用，它响应不同的碳源而改变其在细胞核和内质网膜之间的定位。此外，对 YAS1、YAS2 和 YAS3 缺失突变体中 12 个 ALK 基因的定量实时 PCR 分析表明，Yas3p 参与多种 ALK 基因（包括 ALK1）的转录抑制。相比之下，YAS3 缺失并不影响解脂耶氏酵母中 INO1 直向同源物的 mRNA 水平，表明 Opi1 家族转录因子的功能多样性。

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Yas3p, an Opi1 family transcription factor, regulates cytochrome P450 expression in response to n-alkanes in Yarrowia lipolytica.

In the alkane-assimilating yeast Yarrowia lipolytica, the expression of ALK1, a gene encoding cytochrome P450 that catalyzes the first step of n-alkane oxidation, is induced by n-alkanes. We previously demonstrated that two basic helix-loop-helix proteins, Yas1p and Yas2p, activate the transcription of ALK1 in an alkane-dependent manner by forming a heterocomplex and binding to alkane-responsive element 1 (ARE1), a cis-acting element in the ALK1 promoter. Here we identified an Opi1 family transcription factor, Yas3p, involved in the alkane-dependent transcription regulation of ALK genes. Deletion of YAS3 caused a significant increase in ALK1 mRNA in cells grown on glucose, glycerol, and n-alkanes. The YAS3 deletion also resulted in a marked elevation of reporter gene expression driven by an ARE1-containing promoter on glycerol and n-decane. Bacterially expressed Yas3p bound specifically to Yas2p, but not to Yas1p, in vitro. In addition, although green fluorescent protein-tagged Yas3p was localized in the nucleus in glucose-containing medium, it changed its localization to an endoplasmic reticulum-like compartment upon transfer to medium containing n-decane. These findings suggest that Yas3p functions as a master regulator of transcriptional response, which changes its localization between the nucleus and endoplasmic reticulum membrane in response to different carbon sources. Furthermore, quantitative real time PCR analysis of 12 ALK genes in YAS1, YAS2, and YAS3 deletion mutants suggested that Yas3p is involved in the transcriptional repression of a variety of ALK genes, including ALK1. In contrast, YAS3 deletion did not affect the mRNA level of an INO1 ortholog in Y. lipolytica, indicating functional diversity of Opi1 family transcription factors.

更新日期：2017-01-31

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