活跃的人类组蛋白前 mRNA 3'端加工机制的结构。
Structure of an active human histone pre-mRNA 3'-end processing machinery.
机构信息
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Laboratory of Molecular Electron Microscopy, Rockefeller University, New York, NY 10065, USA.
出版信息
Science. 2020 Feb 7;367(6478):700-703. doi: 10.1126/science.aaz7758.
Abstract
The 3'-end processing machinery for metazoan replication-dependent histone precursor messenger RNAs (pre-mRNAs) contains the U7 small nuclear ribonucleoprotein and shares the key cleavage module with the canonical cleavage and polyadenylation machinery. We reconstituted an active human histone pre-mRNA processing machinery using 13 recombinant proteins and two RNAs and determined its structure by cryo-electron microscopy. The overall structure is highly asymmetrical and resembles an amphora with one long handle. We captured the pre-mRNA in the active site of the endonuclease, the 73-kilodalton subunit of the cleavage and polyadenylation specificity factor, poised for cleavage. The endonuclease and the entire cleavage module undergo extensive rearrangements for activation, triggered through the recognition of the duplex between the authentic pre-mRNA and U7 small nuclear RNA (snRNA). Our study also has notable implications for understanding canonical and snRNA 3'-end processing.
摘要
真核生物复制依赖性组蛋白前体信使 RNA（pre-mRNA）的 3'-末端加工机制包含 U7 小核核糖核蛋白，并与典型的切割和多聚腺苷酸化机制共享关键的切割模块。我们使用 13 种重组蛋白和两种 RNA 重新组装了活性的人组蛋白 pre-mRNA 加工机制，并通过冷冻电子显微镜确定了其结构。整体结构高度不对称，类似于带有长柄的 Amphora。我们将 pre-mRNA 捕获在内切酶的活性位点中，即切割和多聚腺苷酸化特异性因子的 73 千道尔顿亚基，准备进行切割。内切酶和整个切割模块发生广泛的重排以被激活，这是通过识别真实的 pre-mRNA 和 U7 小核 RNA（snRNA）之间的双链体触发的。我们的研究对于理解典型和 snRNA 3'-末端加工也具有重要意义。
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