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Arabidopsis FLL2 promotes liquid-liquid phase separation of polyadenylation complexes

机译：拟南芥FLL2促进多腺苷酸化合物的液 - 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：细胞生物化学的一个重要组成部分是非膜质隔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。在体外已经过度研究了液液相分离所需的多价相互作用（1,3）。然而，在体内调节该过程是较差的。在这里，我们通过拟南芥RNA结合蛋白FCA功能所需的遗传筛网靶向因子鉴定液体液相分离的体内调节剂。 FCA含有朊病毒域，其在体外相分开，并且在体内表现出与相分离一致的行为。突变筛网鉴定了FCA核体形成的FLL2，卷曲卷蛋白的功能要求。 FCA通过在拟南芥基因组（3,4）中的许多位点上促进近端多腺苷酸来减少转录读数。需要FLL2以促进该近端多腺苷酸，但不是FCA对靶标RNA的结合。 FLL2的异位表达增加了FCA核体的尺寸和数量。用甲醛的交联在FLL2，FCA和RNA 3'-终端加工机械的聚合酶和核酸酶模块之间的体内相互作用中捕获。这些3'RNA加工组分与体内核体中的FCA分开化，表明FCA核体分区3'-末端处理因子，以增强特定位点的多苯基化。我们的研究结果表明，卷曲卷材蛋白可以促进液体相分离，这扩大了我们对管理液体样体体内动态的原理的理解。

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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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