Yas3p, an Opi1 Family Transcription Factor, Regulates Cytochrome P450 Expression in Response to n-Alkanes in Yarrowia lipolytica
Yas3p, an Opi1 Family Transcription Factor, Regulates Cytochrome P450 Expression in Response to n-Alkanes in Yarrowia lipolytica
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DOI：
10.1074/jbc.m806864200
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发表时间：
2009-03-13
影响因子：
4.8
通讯作者：
Ohta, Akinori
中科院分区：
文献类型：
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作者：
Hirakawa, Kiyoshi;Kobayashi, Satoshi;Ohta, Akinori
在烷烃同化酵母Yarrowia lipolytica中，正烷烃诱导ALK 1的表达，ALK 1是编码催化正烷烃氧化第一步的细胞色素P450的基因。我们以前证明，两个基本的螺旋-环-螺旋蛋白，Yas 1 p和Yas 2 p，激活转录的ALK 1在一个烷烃依赖的方式，通过形成一个异源复合物和结合到烷烃反应元件1（ARE 1），在ALK 1启动子的顺式作用元件。在这里，我们确定了Opi 1家族的转录因子，Yas 3 p，参与的烷烃依赖性转录调控的ALK基因。YAS 3的缺失导致在葡萄糖、甘油和正烷烃上生长的细胞中ALK 1 mRNA显著增加。YAS 3缺失也导致了由甘油和正癸烷上含ARE 1启动子驱动的报告基因表达的显著升高。在体外，细菌表达的Yas 3 p特异性结合Yas 2 p，但不结合Yas 1 p。此外，虽然绿色荧光蛋白标记的Yas 3 p在含葡萄糖的培养基中定位在细胞核中，但在转移到含正癸烷的培养基中后，它改变了其定位到内质网样隔室。这些发现表明，Yas 3 p作为转录反应的主调节器发挥作用，其响应于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，YAS 1、YAS 2和YAS 3缺失突变体中12个ALK基因的定量真实的时间PCR分析表明，Yas 3 p参与了多种ALK基因（包括ALK 1）的转录抑制。相反，YAS 3缺失不影响Y中INO 1直系同源物的mRNA水平。Lipolytica，表明Opi 1家族转录因子的功能多样性。
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.