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Binding and Regulation of Transcription by Yeast Ste12 Variants To Drive Mating and Invasion Phenotypes.
GENETICS ( IF 3.3 ) Pub Date : 2019-12-06 , DOI: 10.1534/genetics.119.302929
Wei Zhou 1, 2 , Michael W Dorrity 1 , Kerry L Bubb 1 , Christine Queitsch 1 , Stanley Fields 3, 4
Affiliation  

Amino acid substitutions are commonly found in human transcription factors, yet the functional consequences of much of this variation remain unknown, even in well-characterized DNA-binding domains. Here, we examine how six single-amino acid variants in the DNA-binding domain of Ste12-a yeast transcription factor regulating mating and invasion-alter Ste12 genome binding, motif recognition, and gene expression to yield markedly different phenotypes. Using a combination of the "calling-card" method, RNA sequencing, and HT-SELEX (high throughput systematic evolution of ligands by exponential enrichment), we find that variants with dissimilar binding and expression profiles can converge onto similar cellular behaviors. Mating-defective variants led to decreased expression of distinct subsets of genes necessary for mating. Hyper-invasive variants also decreased expression of subsets of genes involved in mating, but increased the expression of other subsets of genes associated with the cellular response to osmotic stress. While single-amino acid changes in the coding region of this transcription factor result in complex regulatory reconfiguration, the major phenotypic consequences for the cell appear to depend on changes in the expression of a small number of genes with related functions.

中文翻译:

酵母 Ste12 变体转录的结合和调节以驱动交配和入侵表型。

氨基酸取代在人类转录因子中很常见,但这种变异的功能后果仍然未知,即使是在已充分表征的 DNA 结合域中也是如此。在这里,我们研究了 Ste12(一种调节交配和入侵的酵母转录因子)DNA 结合域中的 6 个单氨基酸变体如何改变 Ste12 基因组结合、基序识别和基因表达,从而产生明显不同的表型。结合使用“名片”方法、RNA 测序和 HT-SELEX(通过指数富集进行配体的高通量系统进化),我们发现具有不同结合和表达谱的变体可以收敛到相似的细胞行为。交配缺陷变异导致交配所需基因的不同子集的表达减少。高侵入性变异还降低了参与交配的基因子集的表达,但增加了与细胞对渗透应激的反应相关的其他基因子集的表达。虽然该转录因子编码区的单个氨基酸变化会导致复杂的调控重构,但细胞的主要表型后果似乎取决于少数具有相关功能的基因表达的变化。
更新日期:2020-08-22
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