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首页> 美国卫生研究院文献>The EMBO Journal >A novel switch-activating site (SAS1) and its cognate binding factor (SAP1) required for efficient mat1 switching in Schizosaccharomyces pombe.
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A novel switch-activating site (SAS1) and its cognate binding factor (SAP1) required for efficient mat1 switching in Schizosaccharomyces pombe.

机译：一个新型的开关激活位点（SAS1）及其同源结合因子（SAP1）是粟酒裂殖酵母中有效进行mat1转换所必需的。

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摘要

The pattern of parental DNA strand inheritance at the mating type locus (mat1) determines the pattern of mat1 switching in a cell lineage by regulating the formation of the site-specific double-stranded break (DSB) required for mating type interconversion in Schizosaccharomyces pombe. To study the molecular basis of this programmable cell type change, we conducted structural and functional analyses of the DNA sequence flanking the DSB at mat1. We have identified and characterized a DNA-binding activity that interacts with a specific sequence located 140 bp from the DSB site. Deletion analysis of DNA sequences located distal to mat1 cassette revealed the presence of at least two switch-activating sites (SAS1 and SAS2), both of which are required for generating an efficient level of DSBs and consequently, for efficient switching. We found that SAS1 overlaps with the target site of the DNA-binding activity called SAP1 (for switch-activating protein). Point mutations generated in the SAS1 element that adversely affect binding of SAP1 protein in vitro were found to reduce the efficiency of switching in vivo, suggesting the requirement of SAP1 for switching. Pedigree analysis revealed that SAS1 is equally required for initial switching (one switch in four grand-daughters of a cell) and for consecutive switching (where the sister of a recently switched cell switches again), indicating that the two developmentally asymmetric cell divisions required to generate a particular pattern of switching share the same molecular control mechanism.
机译：交配型基因座（mat1）上亲本DNA链遗传的模式通过调节粟酒裂殖酵母交配型相互转化所需的位点特异性双链断裂（DSB）的形成，来确定细胞谱系中mat1转换的模式。为了研究这种可编程细胞类型变化的分子基础，我们对位于mat1的DSB侧翼的DNA序列进行了结构和功能分析。我们已经鉴定并鉴定了一种DNA结合活性，该活性与距DSB位点140 bp的特定序列相互作用。对位于mat1盒远端的DNA序列的缺失分析显示，存在至少两个开关激活位点（SAS1和SAS2），这两个位点对于生成有效水平的DSB并因此对于有效转换都是必需的。我们发现SAS1与称为SAP1的DNA结合活性的靶位点重叠（用于开关激活蛋白）。发现在SAS1元件中产生的点突变对体外SAP1蛋白的结合产生不利影响，从而降低了体内转换的效率，这表明需要SAP1进行转换。谱系分析显示，对于初始切换（一个单元的四个孙女中的一个切换）和连续切换（其中最近切换的单元的姐妹再次切换），均需要SAS1，这表明两个发育不对称的单元划分需要产生特定的开关模式共享相同的分子控制机制。

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