Trm1p, a Zn(II)2Cys6-type transcription factor, is a master regulator of methanol-specific gene activation in the methylotrophic yeast Candida boidinii
Trm1p, a Zn(II)2Cys6-type transcription factor, is a master regulator of methanol-specific gene activation in the methylotrophic yeast Candida boidinii
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DOI：
10.1128/ec.00403-07
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发表时间：
2008-03-01
期刊：
影响因子：
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通讯作者：
Sakai, Yasuyoshi
中科院分区：
文献类型：
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作者：
Sasano, Yu;Yurimoto, Hiroya;Sakai, Yasuyoshi
甲基营养酵母通常被用作外源基因表达的宿主。在这项研究中，我们描述了一个新的基因，TRM1，在假丝酵母中，负责几个甲醇诱导的启动子的转录激活。编码蛋白Trm1p是一种锌(II)(2)半胱氨酸(6)型锌簇蛋白。TRM1的缺失完全抑制了在甲醇上的生长，但在葡萄糖或其他非发酵碳源、甘油、乙醇或油酸上不会导致生长缺陷。Trm1p负责被测试的五个甲醇诱导启动子的转录激活，但不负责过氧化体组装或过氧化体蛋白运输。TRM1基因的表达是结构性的，Trm1p定位于细胞核，与碳源无关。在二羟丙酮合成酶基因的启动子中存在两个顺式作用的甲醇反应元件(MRE)，即MRE1和MRE2。Trm1p被证明是依赖MRE1的甲醇诱导基因表达所必需的。染色质免疫沉淀分析表明，Trm1p在甲醇诱导时与五个甲醇诱导的启动子结合，但在葡萄糖培养的细胞中不结合。因此，TRM1基因编码了一个主要的转录调节因子，负责甲醇营养酵母中甲醇特异性基因的激活。
The methylotrophic yeasts are commonly used as hosts for heterologous gene expression. In this study, we describe a novel gene, TRM1, in Candida boidinii, responsible for the transcriptional activation of several methanol-inducible promoters. The encoded protein, Trm1p, is a Zn(II) (2)Cys(6)-type zinc cluster protein. Deletion of TRM1 completely inhibits growth on methanol but causes no growth defect on glucose or other nonfermentative carbon sources, glycerol, ethanol, or oleate. Trm1p is responsible for transcriptional activation of five methanol-inducible promoters tested, but not for peroxisome assembly or peroxisomal protein transport. Expression of the TRM1 gene was constitutive, and Trm1p localizes to the nuclei regardless of the carbon source. Two cis-acting methanol response elements ( MREs), MRE1 and MRE2 are present in the promoter of the dihydroxyacetone synthase gene. Trm1p is shown to be required for MRE1-dependent methanol inducible gene expression. Chromatin immunoprecipitation assays reveal that Trm1p binds to five methanol-inducible promoters upon methanol induction but does not bind in glucose-grown cells. Thus, the TRM1 gene encodes a master transcriptional regulator responsible for methanol-specific gene activation in the methylotrophic yeasts.