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首页> 外文期刊>Molecular and Cellular Biology >Telomere Formation by Rap1p Binding Site Arrays Reveals End-Specific Length Regulation Requirements and Active Telomeric Recombination
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Telomere Formation by Rap1p Binding Site Arrays Reveals End-Specific Length Regulation Requirements and Active Telomeric Recombination

机译：Rap1p结合位点阵列的端粒形成揭示了末端特定的长度调节要求和主动端粒重组。

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Rap1p, the major telomere repeat binding protein in yeast, has been implicated in both de novo telomere formation and telomere length regulation. To characterize the role of Rap1p in these processes in more detail, we studied the generation of telomeres in vivo from linear DNA substrates containing defined arrays of Rap1p binding sites. Consistent with previous work, our results indicate that synthetic Rap1p binding sites within the internal half of a telomeric array are recognized as an integral part of the telomere complex in an orientation-independent manner that is largely insensitive to the precise spacing between adjacent sites. By extending the lengths of these constructs, we found that several different Rap1p site arrays could never be found at the very distal end of a telomere, even when correctly oriented. Instead, these synthetic arrays were always followed by a short (≈100-bp) “cap” of genuine TG repeat sequence, indicating a remarkably strict sequence requirement for an end-specific function(s) of the telomere. Despite this fact, even misoriented Rap1p site arrays promote telomere formation when they are placed at the distal end of a telomere-healing substrate, provided that at least a single correctly oriented site is present within the array. Surprisingly, these heterogeneous arrays of Rap1p binding sites generate telomeres through a RAD52-dependent fusion resolution reaction that results in an inversion of the original array. Our results provide new insights into the nature of telomere end capping and reveal one way by which recombination can resolve a defect in this process.
机译：Rap1p，酵母中的主要端粒重复结合蛋白，已经参与了从头端粒形成和端粒长度调节。为了更详细地描述Rap1p在这些过程中的作用，我们研究了从包含Rap1p结合位点的确定阵列的线性DNA底物体内生成端粒的过程。与以前的工作一致，我们的结果表明，端粒阵列内部一半内的合成Rap1p结合位点以定向无关的方式被认为是端粒复合物的组成部分，该方式对相邻位点之间的精确间距不敏感。通过延长这些构建体的长度，我们发现即使端粒正确定向，也永远不会在端粒的最远端找到几个不同的Rap1p位点阵列。取而代之的是，这些合成阵列后面总是紧接一个真正的TG重复序列（约100 bp）的“上限”，表明端粒的末端特异功能对序列的要求非常严格。尽管有这个事实，但只要Rap1p位点阵列位于端粒修复基质的远端，即使取向错误的Rap1p位点阵列也要促进端粒的形成，前提是该阵列中至少应存在一个正确取向的位点。令人惊讶的是，这些异质性的Rap1p结合位点阵列通过依赖于 RAD52 的融合分辨反应生成端粒，从而导致原始阵列倒置。我们的结果提供了有关端粒末端封端性质的新见解，并揭示了重组可以解决此过程中缺陷的一种方法。

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