Macromolecular Machines Laboratory, The Francis Crick Institute, NW1 1AT, London, UK.
Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3JR, UK.
Nat Commun. 2019 Sep 13;10(1):4189. doi: 10.1038/s41467-019-12007-w.
Retroviral integrase can efficiently utilise nucleosomes for insertion of the reverse-transcribed viral DNA. In face of the structural constraints imposed by the nucleosomal structure, integrase gains access to the scissile phosphodiester bonds by lifting DNA off the histone octamer at the site of integration. To clarify the mechanism of DNA looping by integrase, we determined a 3.9 Å resolution structure of the prototype foamy virus intasome engaged with a nucleosome core particle. The structural data along with complementary single-molecule Förster resonance energy transfer measurements reveal twisting and sliding of the nucleosomal DNA arm proximal to the integration site. Sliding the nucleosomal DNA by approximately two base pairs along the histone octamer accommodates the necessary DNA lifting from the histone H2A-H2B subunits to allow engagement with the intasome. Thus, retroviral integration into nucleosomes involves the looping-and-sliding mechanism for nucleosomal DNA repositioning, bearing unexpected similarities to chromatin remodelers.
逆转录病毒整合酶可以有效地将逆转录病毒 DNA 插入核小体。面对核小体结构所施加的结构限制,整合酶通过在整合部位将 DNA 从组蛋白八聚体上抬起,从而获得对切口磷酸二酯键的访问。为了阐明整合酶介导的 DNA 环化机制,我们确定了与核小体核心颗粒结合的原型泡沫病毒整合酶复合物的 3.9Å 分辨率结构。结构数据和互补的单分子Förster 共振能量转移测量结果表明,整合部位附近的核小体 DNA 臂发生扭曲和滑动。沿着组蛋白八聚体滑动核小体 DNA 大约两个碱基对,从而使 DNA 从组蛋白 H2A-H2B 亚基上抬起,以允许与整合酶复合物结合。因此,逆转录病毒整合到核小体中涉及核小体 DNA 重定位的环化-滑动机制,这与染色质重塑剂具有出人意料的相似性。