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首页> 外文期刊>Molecular and Cellular Biology >Detection of leucine-independent DNA site occupancy of the yeast Leu3p transcriptional activator in vivo.

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Detection of leucine-independent DNA site occupancy of the yeast Leu3p transcriptional activator in vivo.

机译：体内检测酵母Leu3p转录激活因子的亮氨酸非依赖性DNA位点占有率。

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The product of the Saccharomyces cerevisiae LEU3 gene, Leu3p, is a transcriptional activator which regulates leucine biosynthesis in response to intracellular levels of leucine through the biosynthetic intermediate alpha-isopropylmalate. We devised a novel assay to examine the DNA site occupancy of Leu3p under different growth conditions, using a reporter gene with internal Leu3p-binding sites. Expression of the reporter is inhibited by binding of nuclear Leu3p to these sites; inhibition is dependent on the presence of the sites in the reporter, on the integrity of the Leu3p DNA-binding domain, and, surprisingly, on the presence of a transcriptional activation domain in the inhibiting protein. By this assay, Leu3p was found to occupy its binding site under all conditions tested, including high and low levels of leucine and in the presence and absence of alpha-isopropylmalate. The localization of Leu3p to the nucleus was confirmed by immunofluorescence staining of cells expressing epitope-tagged Leu3p derivatives. We conclude that Leu3p regulates transcription in vivo without changing its intracellular localization and DNA site occupancy.

机译：啤酒酵母LEU3基因的产物Leu3p是一种转录激活因子，通过生物合成中间体α-异丙基苹果酸来响应亮氨酸的细胞内水平，从而调节亮氨酸的生物合成。我们设计了一种新颖的检测方法，使用带有内部Leu3p结合位点的报告基因，在不同的生长条件下检查Leu3p的DNA位点占用。核Leu3p与这些位点的结合可抑制报告基因的表达。抑制作用取决于报告基因中位点的存在，Leu3p DNA结合结构域的完整性，以及令人惊讶地取决于抑制蛋白中转录激活结构域的存在。通过该测定，发现Leu3p在所有测试的条件下都占据其结合位点，包括高和低水平的亮氨酸，以及存在和不存在α-异丙基苹果酸。通过对表达表位标记的Leu3p衍生物的细胞进行免疫荧光染色，证实了Leu3p在细胞核中的定位。我们得出的结论是，Leu3p在体内调节转录，而不改变其细胞内定位和DNA位点占用。

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《Molecular and Cellular Biology》 |1995年第8期|共10页
作者
C R Kirkpatrick; P Schimmel;
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中图分类细胞生物学;
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1. A novel DNA binding motif for yeast zinc cluster proteins: the Leu3p and Pdr3p transcriptional activators recognize everted repeats. [J] . K Hellauer, M H Rochon, B Turcotte Molecular and Cellular Biology . 1996,第11期

机译：酵母锌簇蛋白的新型DNA结合基序：Leu3p和Pdr3p转录激活因子识别外翻重复序列。
2. The yeast SWI-SNF complex facilitates binding of a transcriptional activator to nucleosomal sites in vivo. [J] . L G Burns, C L Peterson Molecular and Cellular Biology . 1997,第8期

机译：酵母SWI-SNF复合物促进转录激活剂与体内核小体位点的结合。
3. The monomer of the yeast transcriptional activator GCN4 recognizes its dimer binding DNA target sites without dimerization [J] . Cao W., Klein DE., Wei LY., Thermochimica Acta: An International Journal Concerned with the Broader Aspects of Thermochemistry and Its Applications to Chemical Problems . 2000,第1期

机译：酵母转录激活因子GCN4的单体识别其二聚体结合DNA靶位点而无需二聚化
4. Mining Yeast Transcriptional Regulatory Modules from Factor DNA-Binding Sites and Gene Expression Data [C] . Tho Hoan Pham, Kenji Satou, Tu Bao Ho International Workshop on Bioinformatics and Systems Biology . 2004

机译：从因子DNA结合位点和基因表达数据的矿床转录调节模块
5. Self-silencing of yeast transcriptional activator Leu3p: Studies on mechanism, specificity and regulation of the activation domains masking process. [D] . Wang, Dake. 1998

机译：酵母转录激活因子Leu3p的自我沉默：对激活域掩盖过程的机制，特异性和调控的研究。
6. Detection of leucine-independent DNA site occupancy of the yeast Leu3p transcriptional activator in vivo. [O] . C R Kirkpatrick, P Schimmel 1995

机译：体内检测酵母Leu3p转录激活因子的亮氨酸非依赖性DNA位点占有率。
7. Detection of leucine-independent DNA site occupancy of the yeast Leu3p transcriptional activator in vivo. [O] . Kirkpatrick, C R, Schimmel, P 1995

机译：体内检测酵母Leu3p转录激活因子的亮氨酸非依赖性DNA位点占有率。

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