GENERATING YEAST TRANSCRIPTIONAL ACTIVATORS CONTAINING NO YEAST PROTEIN SEQUENCES
GENERATING YEAST TRANSCRIPTIONAL ACTIVATORS CONTAINING NO YEAST PROTEIN SEQUENCES
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DOI：
10.1038/350250a0
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发表时间：
1991-03-21
期刊：
影响因子：
64.8
通讯作者：
PTASHNE, M
中科院分区：
文献类型：
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作者：
RUDEN, DM;MA, J;PTASHNE, M
我们先前报道，当与GAL 4（1-147）融合时，由大肠杆菌基因组DNA片段编码的大约1%的短肽在酵母中充当转录激活区，GAL 4（1-147）是酵母转录激活因子GAL 4的DNA结合部分（参考文献1）。 Struhl质疑我们已经鉴定出在没有酵母转录激活序列的情况下发挥作用的新的转录激活序列的结论2。 他的批评基于两个考虑：第一，GAL 4（1-147）含有一个酸性片段（随后的实验表明，该区域含有一个体外弱激活区3）;第二，当细菌阻遏物的DNA结合结构域莱克萨（1-87）被用作DNA结合单元2时，分离新激活区的尝试失败了。 我们在此报告了我们原始实验的重复，使用完整的莱克萨分子莱克萨（1-202）作为DNA结合区，而不是GAL 4（1-147）或莱克萨（1-87）。 我们发现，与最初的实验一样，大约1%的由E. coli基因组片段与完整的莱克萨融合后可作为转录激活区。 我们确定其序列的所有新的激活区都含有过量的酸性氨基酸（见表1）。
WE previously reported that roughly 1% of the short peptides encoded by Escherichia coli genomic DNA fragments act as transcriptional activating regions in yeast when fused to GAL4(1-147), a DNA-binding portion of the yeast transcriptional activator GAL4 (ref. 1). Struhl questioned the conclusion that we had identified new transcriptional activating sequences that function in the absence of yeast transcriptional activating sequences 2. His criticism was based on two considerations: first, GAL4(1-147) contains an acidic segment (and subsequent experiments have shown that this region contains a weak activating region in vitro 3); second, attempts to isolate new activating regions failed when the DNA-binding domain of a bacterial repressor, LexA(1-87), was used as the DNA-binding unit 2. We report here a repeat of our original experiment using the complete LexA molecule LexA(1-202) as the DNA-binding region, instead of GAL4(1-147) or LexA(1-87). We find that, as in the original experiment, about 1% of the short peptides encoded by E. coli genomic fragments act as transcriptional activating regions when fused to intact LexA. All of the new activating regions whose sequences we determined bore an excess of acidic amino acids (see Table 1).