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DNA self-recognition in the structure of Pot1 bound to telomeric single-stranded DNA

机译：与端粒单链DNA结合的Pot1结构中的DNA自我识别

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摘要

Telomeres, specialized protein-DNA complexes that cap the ends of linear chromosomes, are essential for protecting chromosomes from degradation and end-to-end fusions. The Potl (protection of telomeres 1) protein is a widely distributed eukaryotic end-capping protein, having been identified in fission yeast, microsporidia, plants and animals. Schizosaccharomyces pombe Pot1p is essential for telomere maintenance, and human POT1 has been implicated in telomerase regulation. Potl binds telomeric single-stranded DNA (ssDNA) with exceptionally high sequence specificity, the molecular basis of which has been unknown. Here we describe the 1.9-A-resolution crystal structure of the ammo-terminal DNA-binding domain of S. pombe Potlp complexed with ssDNA. The protein adopts an oligonu-cleotide/oligosaccharide-binding (OB) fold with two loops that protrude to form a clamp for ssDNA binding. The structure explains the sequence specificity of binding: in the context of the Potl protein, DNA self-recognition involving base-stacking and unusual G-T base pairs compacts the DNA. Any sequence change disrupts the ability of the DNA to form this structure, preventing it from contacting the array of protein hydrogen-bonding groups. The structure also explains how Potlp avoids binding the vast excess of RNA in the nucleus.
机译：端粒是一种专门的蛋白质-DNA复合物，可覆盖线性染色体的末端，对于保护染色体免受降解和端对端融合至关重要。 Potl（端粒1的保护）蛋白是一种广泛分布的真核末端封端蛋白，已在裂变酵母，微孢子虫，植物和动物中鉴定出。粟酒裂殖酵母Pot1p对端粒的维持至关重要，人类POT1已参与端粒酶的调控。 Potl以异常高的序列特异性结合端粒单链DNA（ssDNA），其分子基础尚不清楚。在这里，我们描述了与ssDNA复合的粟酒裂殖酵母的弹药末端DNA结合域的1.9-A分辨率晶体结构。该蛋白采用寡核苷酸-寡核苷酸/寡糖结合（OB）折叠，带有两个环，突出形成用于ssDNA结合的夹具。该结构解释了结合的序列特异性：在Potl蛋白的背景下，涉及碱基堆积和异常G-T碱基对的DNA自我识别使DNA紧密结合。任何序列变化都会破坏DNA形成此结构的能力，从而阻止其与蛋白质氢键基团阵列接触。该结构还解释了Potlp如何避免结合细胞核中大量过量的RNA。

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  • 来源
    《Nature》 |2003年第6963期|p.198-203|共6页
  • 作者

    Ming Lei; Elaine R. Podell; Peter Baumann; Thomas R. Cech;

  • 作者单位

    Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309-0215, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 自然科学总论;
  • 关键词

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