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首页> 外文期刊>Nucleic Acids Research >Resolution of the Holliday junction recombination intermediate by human GEN1 at the single-molecule level
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Resolution of the Holliday junction recombination intermediate by human GEN1 at the single-molecule level

机译：通过人Gen1在单分子水平上通过人Gen1分辨Holliday结重组的分辨率

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摘要

Human GEN1 is a cytosolic homologous recombination protein that resolves persisting four-way Holliday junctions (HJ) after the dissolution of the nuclear membrane. GEN1 dimerization has been suggested to play key role in the resolution of the HJ, but the kinetic details of its reaction remained elusive. Here, single-molecule FRET shows how human GEN1 binds the HJ and always ensures its resolution within the lifetime of the GEN1-HJ complex. GEN1 monomer generally follows the isomer bias of the HJ in its initial binding and subsequently distorts it for catalysis. GEN1 monomer remains tightly bound with no apparent dissociation until GEN1 dimer is formed and the HJ is fully resolved. Fast on- and slow off-rates of GEN1 dimer and its increased affinity to the singly-cleaved HJ enforce the forward reaction. Furthermore, GEN1 monomer binds singly-cleaved HJ tighter than intact HJ providing a fail-safe mechanism if GEN1 dimer or one of its monomers dissociates after the first cleavage. The tight binding of GEN1 monomer to intact- and singly-cleaved HJ empowers it as the last resort to process HJs that escape the primary mechanisms.
机译：人Gen1是胞质同源重组蛋白，其在核膜溶解后持久地持续四向Holliday联结（HJ）。已经提出了Gen1二聚体在HJ的分辨率中起着关键作用，但其反应的动力学细节仍然难以捉摸。这里，单分子FRET显示人Gen1如何结合HJ并始终确保其在Gen1-HJ复合物的寿命范围内的分辨率。 Gen1单体通常在其初始结合中遵循HJ的异构体偏压，随后将其扭曲以催化。 Gen1单体保持紧密结合，直至形成Gen1二聚体并完全分解HJ。 Gen1二聚体的快速和慢速速率及其对单裂纹HJ的增加的亲和力强制执行前进反应。此外，Gen1单体与完整的HJ相比单独切割的Hj，如果Gen1二聚体或其单体在第一次切割后解离，则不能提供故障安全机制。 Gen1单体对完整和单独切割的HJ的紧密结合赋予它作为逃离主要机制的HJ的最后一个手段。

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  • 来源
    《Nucleic Acids Research》 |2019年第4期|共15页
  • 作者

    Sobhy Mohamed A.; Brailc Amer; Raducanu Vlad-Stefan; Takahashi Masateru; Tehseen Muhammad; Rashid Fahad; Zaher Manal S.; Hamdan Samir M.;

  • 作者单位

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

    King Abdullah Univ Sci &

    Technol Biol &

    Environm Sci &

    Engn Div Lab DNA Replicat &

    Recombinat Thuwal 239556900 Saudi Arabia;

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  • 正文语种 eng
  • 中图分类 生物化学;
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