<br>嗜热热菌 HB8 转录因子的一般和基因组 DNA 结合特异性 TTHB023。,Biomolecules

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嗜热热菌 HB8 转录因子的一般和基因组 DNA 结合特异性 TTHB023。
Biomolecules ( IF 4.8 ) Pub Date : 2020-01-06 , DOI: 10.3390/biom10010094
James Shell Cox _{1,

2} , Michael W Van Dyke ₁

Affiliation

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 序列为了解它们调节的基因集以及由此了解它们的潜在生物学功能提供了重要线索。可以通过同源搜索和遗传手段获得见解。然而，这些方法可能具有误导性，尤其是在比较远亲生物体或复杂转录调控的情况下。在这项工作中，我们使用基于生物化学的方法来确定与嗜热菌 HB8 TetR 家族转录因子特异性结合的 DNA 序列谱TTHB023。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 TTHB023 具有纳摩尔结合亲和力。分析嗜热 T 菌 HB8 基因组，将几个TTHB023共有结合位点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出来，表面上是因为它们可能受到嗜热热菌 HB8 TetR 家族转录阻遏蛋白TTHA0167的共同调节。我们的调查提供了额外的证据，支持基于生物化学的方法对表征假定转录因子的有用性，尤其是在合作调节的情况下。

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General and Genomic DNA-Binding Specificity for the Thermus thermophilus HB8 Transcription Factor TTHB023.

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.

更新日期：2020-01-06

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