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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Gated rotation mechanism of site-specific recombination by φC31 integrase
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Gated rotation mechanism of site-specific recombination by φC31 integrase

机译：通过φC31整合酶进行位点重组的门控旋转机制

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摘要

Integrases, such as that of the Streptomyces temperate bacterio-phage φC31, promote site-specific recombination between DNA sequences in the bacteriophage and bacterial genomes to integrate or excise the phage DNA. φC31 integrase belongs to the serine recombinase family, a large group of structurally related enzymes with diverse biological functions. It has been proposed that serine integrases use a "subunit rotation" mechanism to exchange DNA strands after double-strand DNA cleavage at the two recombining art sites, and that many rounds of subunit rotation can occur before the strands are religated. We have analyzed the mechanism of φC31 integrase-mediated recombination in a topologically constrained experimental system using hybrid "phes" recombination sites, each of which comprises a φC31 att site positioned adjacent to a regulatory sequence recognized by Tn3 resolvase. The topologies of reaction products from circular substrates containing two phes sites support a right-handed subunit rotation mechanism for catalysis of both integrative and excisive recombination. Strand exchange usually terminates after a single round of 180° rotation. However, multiple processive "360° rotation" rounds of strand exchange can be observed, if the recombining sites have nonidentical base pairs at their centers. We propose that a regulatory "gating" mechanism normally blocks multiple rounds of strand exchange and triggers product release after a single round.
机译：诸如温链链霉菌噬菌体φC31的整合体可促进噬菌体中DNA序列与细菌基因组之间的位点特异性重组，从而整合或切除噬菌体DNA。 φC31整合酶属于丝氨酸重组酶家族，是一大类具有多种生物学功能的结构相关酶。已经提出丝氨酸整合使用“亚基旋转”机制在两个重组技术位点双链DNA切割后交换DNA链，并且在链重新连接之前可以发生许多轮亚基旋转。我们使用混合的“ phes”重组位点在拓扑受限的实验系统中分析了φC31整合酶介导的重组的机制，每个位点均包含一个φC31att位点，该位点邻近Tn3溶解酶识别的调控序列。来自包含两个phes位点的圆形底物的反应产物的拓扑结构支持用于整合重组和激发重组的右旋亚基旋转机制。通常在旋转180度后，链交换才会终止。但是，如果重组位点的中心具有不相同的碱基对，则可以观察到多个连续的“ 360°旋转”轮交换。我们建议，监管“门控”机制通常会阻止多轮链交换并在单轮后触发产品释放。

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    Femi J. Olorunniji; Dorothy E. Buck; Sean D. Colloms; Andrew R. McEwan; Margaret C. M. Smith; W. Marshall Stark; Susan J. Rosser;

  • 作者单位

    Institute of Molecular, Cell, and Systems Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom;

    Department of Mathematics, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom;

    Institute of Molecular, Cell, and Systems Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom;

    Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, Scotland, United Kingdom;

    Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, Scotland, United Kingdom,Department of Biology, University of York, York YO10 5DD, United Kingdom;

    Institute of Molecular, Cell, and Systems Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom;

    Institute of Molecular, Cell, and Systems Biology, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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