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Arabidopsis FLL2 promotes liquid-liquid phase separation of polyadenylation complexes

机译：拟南芥FLL2促进多腺苷酸化复合物的液相分离

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摘要

An important component of cellular biochemistry is the concentration of proteins and nucleic acids in non-membranous compartments(1,2). These biomolecular condensates are formed from processes that include liquid-liquid phase separation. The multivalent interactions necessary for liquid-liquid phase separation have been extensively studied in vitro(1,3). However, the regulation of this process in vivo is poorly understood. Here we identify an in vivo regulator of liquid-liquid phase separation through a genetic screen targeting factors required for Arabidopsis RNA-binding protein FCA function. FCA contains prion-like domains that phase-separate in vitro, and exhibits behaviour in vivo that is consistent with phase separation. The mutant screen identified a functional requirement for FLL2, a coiled-coil protein, in the formation of FCA nuclear bodies. FCA reduces transcriptional read-through by promoting proximal polyadenylation at many sites in the Arabidopsis genome(3,4). FLL2 was required to promote this proximal polyadenylation, but not the binding of FCA to target RNA. Ectopic expression of FLL2 increased the size and number of FCA nuclear bodies. Crosslinking with formaldehyde captured in vivo interactions between FLL2, FCA and the polymerase and nuclease modules of the RNA 3'-end processing machinery. These 3' RNA-processing components colocalized with FCA in the nuclear bodies in vivo, which indicates that FCA nuclear bodies compartmentalize 3'-end processing factors to enhance polyadenylation at specific sites. Our findings show that coiled-coil proteins can promote liquid-liquid phase separation, which expands our understanding of the principles that govern the in vivo dynamics of liquid-like bodies.
机译：细胞生物化学的重要组成部分是非膜区室中蛋白质和核酸的浓度（1,2）。这些生物分子缩合物由包括液-液相分离的方法形成。液-液相分离所必需的多价相互作用已在体外进行了广泛研究（1,3）。但是，对该过程在体内的调节知之甚少。在这里，我们通过拟南芥属RNA结合蛋白FCA功能所需的基因筛选，确定了通过液相筛查的液-液相分离的体内调节剂。 FCA包含在体外发生相分离的that病毒样结构域，并在体内表现出与相分离一致的行为。突变体筛选确定了在FCA核小体形成中对盘绕蛋白FLL2的功能需求。 FCA通过促进拟南芥基因组中许多位点的近端聚腺苷酸化来减少转录通透性（3,4）。需要FLL2来促进这种近端聚腺苷酸化，但不能促进FCA与靶RNA的结合。 FLL2的异位表达增加了FCA核体的大小和数量。与甲醛的交联捕获了FLL2，FCA与RNA 3'末端加工机器的聚合酶和核酸酶模块之间的体内相互作用。这些3'RNA加工成分与FCA在体内的核体内共定位，这表明FCA核体将3'末端加工因子区分开以增强特定部位的聚腺苷酸化。我们的发现表明，卷曲螺旋蛋白可以促进液-液相分离，从而扩大了我们对支配液体状体内动力学的原理的理解。

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  • 来源
    《Nature》 |2019年第7755期|265-269|共5页
  • 作者

    Fang Xiaofeng; Wang Liang; Ishikawa Ryo; Li Yaoxi; Fiedler Marc; Liu Fuquan; Calder Grant; Rowan Beth; Weigel Detlef; Li Pilong; Dean Caroline;

  • 作者单位

    John Innes Ctr, Norwich, Norfolk, England;

    Tsinghua Univ, Beijing Adv Innovat Ctr Struct Biol, Tsinghua Univ Peking Univ Joint Ctr Life Sci, Sch Life Sci, Beijing, Peoples R China;

    John Innes Ctr, Norwich, Norfolk, England|Kobe Univ, Grad Sch Agr Sci, Kobe, Hyogo, Japan;

    John Innes Ctr, Norwich, Norfolk, England;

    MRC Lab Mol Biol, Cambridge, England;

    John Innes Ctr, Norwich, Norfolk, England|Queens Univ Belfast, Inst Global Food Secur, Sch Biol Sci, Belfast, Antrim, North Ireland;

    John Innes Ctr, Norwich, Norfolk, England;

    Max Planck Inst Dev Biol, Dept Mol Biol, Tubingen, Germany;

    Max Planck Inst Dev Biol, Dept Mol Biol, Tubingen, Germany;

    Tsinghua Univ, Beijing Adv Innovat Ctr Struct Biol, Tsinghua Univ Peking Univ Joint Ctr Life Sci, Sch Life Sci, Beijing, Peoples R China;

    John Innes Ctr, Norwich, Norfolk, England;

  • 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);
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