General and Genomic DNA-Binding Specificity for the Thermus thermophilus HB8 Transcription Factor TTHB023
General and Genomic DNA-Binding Specificity for the Thermus thermophilus HB8 Transcription Factor TTHB023
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DOI：
10.3390/biom10010094
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发表时间：
2020-01-01
期刊：
影响因子：
5.5
通讯作者：
Van Dyke, Michael W.
中科院分区：
文献类型：
--
作者：
Cox, James Shell;Van Dyke, Michael W.
转录因子是识别特定DNA序列并影响局部转录过程的蛋白质。它们是所有生物体控制特定基因表达的主要手段。了解特定转录因子识别哪些DNA序列为它们调节的基因组以及它们的潜在生物学功能提供了重要线索。可以通过同源性搜索和遗传学手段获得见解。然而，这些方法可能会误导，特别是在比较远亲生物或复杂的转录调控的情况下。在这项工作中，我们使用了一种基于生物化学的方法，以确定光谱的DNA序列特异性结合的嗜热栖热菌HB 8 TetR家族转录因子TTHB 023。发现共有序列5 '-（a/c）Y（g/t）A（A/C）YGryCR（g/t）T（c/a）R（g/t）-3 '与TTHB 023具有纳摩尔结合亲和力。分析了T.在嗜热菌HB 8基因组中，几个TTHB 023共有结合位点被定位到参与脂肪酸生物合成的基因的启动子。值得注意的是，其中一些以前没有通过遗传方法鉴定，表面上是因为它们可能受到嗜热栖热菌HB 8 TetR家族转录抑制子TTHA 0167的共同调节。我们的调查提供了额外的证据支持有用的生物化学为基础的方法来表征假定的转录因子，特别是在合作监管的情况下。
Transcription factors are proteins that recognize specific DNA sequences and affect local transcriptional processes. They are the primary means by which all organisms control specific gene expression. Understanding which DNA sequences a particular transcription factor recognizes provides important clues into the set of genes that they regulate and, through this, their potential biological functions. Insights may be gained through homology searches and genetic means. However, these approaches can be misleading, especially when comparing distantly related organisms or in cases of complicated transcriptional regulation. In this work, we used a biochemistry-based approach to determine the spectrum of DNA sequences specifically bound by the Thermus thermophilus HB8 TetR-family transcription factor TTHB023. The consensus sequence 5 '-(a/c)Y(g/t)A(A/C)YGryCR(g/t)T(c/a)R(g/t)-3 ' was found to have a nanomolar binding affinity with TTHB023. Analyzing the T. thermophilus HB8 genome, several TTHB023 consensus binding sites were mapped to the promoters of genes involved in fatty acid biosynthesis. Notably, some of these were not identified previously through genetic approaches, ostensibly given their potential co-regulation by the Thermus thermophilus HB8 TetR-family transcriptional repressor TTHA0167. Our investigation provides additional evidence supporting the usefulness of a biochemistry-based approach for characterizing putative transcription factors, especially in the case of cooperative regulation.