Linking DNA-binding proteins to their recognition sequences by using protein microarrays
Linking DNA-binding proteins to their recognition sequences by using protein microarrays
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DOI：
10.1073/pnas.0509185103
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发表时间：
2006-06-27
影响因子：
11.1
通讯作者：
Snyder, Michael
中科院分区：
文献类型：
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作者：
Ho, Su-Wen;Jona, Ghil;Snyder, Michael
对全基因组序列和实验数据集的分析揭示了大量的DNA序列基序，这些基序在许多物种中是保守的，并且可能是功能性的。然而，缺乏足够规模的方法来探索这些因素的作用。我们描述了使用蛋白质阵列来识别与感兴趣的DNA序列结合的蛋白质。一个微阵列的282个已知的和潜在的酵母转录因子的产生和探测与寡核苷酸的进化保守序列是潜在的功能。鉴定了与特定DNA序列结合的转录因子。一个以前未表征的DNA结合蛋白，YjI 103，其特征在于详细。我们确定了这种蛋白的结合位点，并确定了一些靶基因，其中许多涉及应激反应和氧化磷酸化。蛋白质微阵列为确定DNA-蛋白质相互作用提供了高通量方法。
Analyses of whole-genome sequences and experimental data sets have revealed a large number of DNA sequence motifs that are conserved in many species and may be functional. However, methods of sufficient scale to explore the roles of these elements are lacking. We describe the use of protein arrays to identify proteins that bind to DNA sequences of interest. A microarray of 282 known and potential yeast transcription factors was produced and probed with oligonucleotides of evolutionarily conserved sequences that are potentially functional. Transcription factors that bound to specific DNA sequences were identified. One previously uncharacterized DNA-binding protein, YjI103, was characterized in detail. We defined the binding site for this protein and identified a number of its target genes, many of which are involved in stress response and oxidative phosphorylation. Protein microarrays offer a high-throughput method for determining DNA-protein interactions.