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Arabidopsis FLL2 promotes liquid–liquid phase separation of polyadenylation complexes
Nature ( IF 50.5 ) Pub Date : 2019-05-01 , DOI: 10.1038/s41586-019-1165-8 Xiaofeng Fang 1 , Liang Wang 2 , Ryo Ishikawa 1, 3 , Yaoxi Li 1 , Marc Fiedler 4 , Fuquan Liu 1, 5 , Grant Calder 1 , Beth Rowan 6 , Detlef Weigel 6 , Pilong Li 2 , Caroline Dean 1
Nature ( IF 50.5 ) Pub Date : 2019-05-01 , DOI: 10.1038/s41586-019-1165-8 Xiaofeng Fang 1 , Liang Wang 2 , Ryo Ishikawa 1, 3 , Yaoxi Li 1 , Marc Fiedler 4 , Fuquan Liu 1, 5 , Grant Calder 1 , Beth Rowan 6 , Detlef Weigel 6 , Pilong Li 2 , Caroline Dean 1
Affiliation
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An important component of cellular biochemistry is the concentration of proteins and nucleic acids in non-membranous compartments1,2. 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 vitro1,3. 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 genome3,4. 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.A genetic screen for factors required by the Arabidopsis RNA-binding protein FCA identifies FLL2 as necessary in the formation of FCA nuclear bodies, and thus a role for FLL2 in liquid–liquid phase separation.
中文翻译：
拟南芥 FLL2 促进聚腺苷酸化复合物的液-液相分离
细胞生物化学的一个重要组成部分是非膜区室 1,2 中蛋白质和核酸的浓度。这些生物分子凝聚物是由包括液-液相分离在内的过程形成的。液-液相分离所需的多价相互作用已在体外进行了广泛研究1,3。然而，对体内这一过程的调节知之甚少。在这里，我们通过拟南芥 RNA 结合蛋白 FCA 功能所需的靶向因子的遗传筛选鉴定了液-液相分离的体内调节剂。FCA 包含在体外相分离的朊病毒样结构域，并在体内表现出与相分离一致的行为。突变筛选确定了 FLL2（一种卷曲螺旋蛋白）在 FCA 核体形成中的功能要求。FCA 通过促进拟南芥基因组中许多位点的近端多聚腺苷酸化来减少转录通读 3,4。FLL2 需要促进这种近端多聚腺苷酸化，而不是促进 FCA 与靶标 RNA 的结合。FLL2 的异位表达增加了 FCA 核体的大小和数量。与甲醛交联捕获 FLL2、FCA 和 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 加工成分在体内与 FCA 共定位于核体中，这表明 FCA 核体将 3'-末端加工因子区分开来以增强特定位点的聚腺苷酸化。我们的研究结果表明，卷曲螺旋蛋白可以促进液-液相分离，
更新日期：2019-05-01
中文翻译：
拟南芥 FLL2 促进聚腺苷酸化复合物的液-液相分离
细胞生物化学的一个重要组成部分是非膜区室 1,2 中蛋白质和核酸的浓度。这些生物分子凝聚物是由包括液-液相分离在内的过程形成的。液-液相分离所需的多价相互作用已在体外进行了广泛研究1,3。然而，对体内这一过程的调节知之甚少。在这里，我们通过拟南芥 RNA 结合蛋白 FCA 功能所需的靶向因子的遗传筛选鉴定了液-液相分离的体内调节剂。FCA 包含在体外相分离的朊病毒样结构域，并在体内表现出与相分离一致的行为。突变筛选确定了 FLL2（一种卷曲螺旋蛋白）在 FCA 核体形成中的功能要求。FCA 通过促进拟南芥基因组中许多位点的近端多聚腺苷酸化来减少转录通读 3,4。FLL2 需要促进这种近端多聚腺苷酸化，而不是促进 FCA 与靶标 RNA 的结合。FLL2 的异位表达增加了 FCA 核体的大小和数量。与甲醛交联捕获 FLL2、FCA 和 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 加工成分在体内与 FCA 共定位于核体中，这表明 FCA 核体将 3'-末端加工因子区分开来以增强特定位点的聚腺苷酸化。我们的研究结果表明，卷曲螺旋蛋白可以促进液-液相分离，
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