Phase separation and molecular ordering of the prion-like domain of the Arabidopsis thermosensory protein EARLY FLOWERING 3.
Phase separation and molecular ordering of the prion-like domain of the Arabidopsis thermosensory protein EARLY FLOWERING 3.
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拟南芥热感应蛋白早期开花3的prion样结构域的相分离和分子排序3。
DOI：
10.1073/pnas.2304714120
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发表时间：
2023-07-11
影响因子：
11.1
通讯作者：
Zubieta, Chloe
中科院分区：
文献类型：
--
作者：
Hutin, Stephanie;Kumita, Janet R.;Strotmann, Vivien I.;Dolata, Anika;Ling, Wai Li;Louafi, Nessim;Popov, Anton;Milhiet, Pierre- Emmanuel;Blackledge, Martin;Nanao, Max H.;Wigge, Philip A.;Stahl, Yvonne;Costa, Luca;Tully, Mark D.;Zubieta, Chloe
关键词：
蛋白质EARLY FLOWERING 3（ELF3）的相分离是温度的函数，由其类朊病毒结构域（PrLD）驱动。在此，我们利用互补的结构、生物物理和显微技术确定相分离的动力学和分子基础。我们证明，在稀相时，ELF3的PrLD形成一种高阶单分散寡聚体，这对相分离至关重要。温度变化触发凝聚相的形成，该凝聚相显示出分子有序性增加。随后，液态凝聚态老化形成水凝胶，基于小角X射线散射以及原子力和电子显微镜呈现出分子堆叠。这些结果解析了一种关键植物温度传感器的不同物理状态的结构。
液 - 液相分离（LLPS）是一种使大分子（包括蛋白质和核酸等复杂聚合物）实现动态区室化的重要机制，并且是物理化学环境的函数。在模式植物拟南芥中，蛋白质EARLY FLOWERING3（ELF3）的LLPS以温度敏感的方式发生，并控制热响应性生长。ELF3包含一个很大程度上无结构的类朊病毒结构域（PrLD），其在体内和体外都是LLPS的驱动因素。PrLD包含一个多聚谷氨酰胺（polyQ）序列，其长度在不同的拟南芥自然品系中有所不同。在此，我们结合生物化学、生物物理和结构技术来研究具有不同polyQ长度的ELF3 PrLD的稀相和凝聚相。我们证明ELF3 PrLD的稀相形成一种不依赖于polyQ序列存在的单分散高阶寡聚体。这种物质以对pH和温度敏感的方式经历LLPS，并且蛋白质的polyQ区域调节相分离的初始阶段。液相迅速老化并形成水凝胶，如荧光和原子力显微镜所示。此外，我们证明水凝胶呈现出一种半有序结构，这是通过小角X射线散射、电子显微镜和X射线衍射确定的。这些实验展示了一种PrLD蛋白质丰富的结构景观，并为描述生物分子凝聚物的结构和生物物理特性提供了一个框架。
Phase separation of the protein EARLY FLOWERING 3 (ELF3) occurs as a function of temperature and is driven by its prion-like domain (PrLD). Here, we determine the dynamics and molecular basis for phase separation using complementary structural, biophysical, and microscopy techniques. We demonstrate that in the dilute phase, the PrLD of ELF3 forms a higher-order monodisperse oligomer, which is vital for phase separation. Changes in temperature trigger the formation of a condensed phase that shows increased molecular ordering. Subsequently, the liquid condensed state ages into a hydrogel, exhibiting molecular stacking based on small-angle X-ray scattering and atomic force and electron microscopy. These results resolve the structure of different physical states for a key plant thermosensor. Liquid–liquid phase separation (LLPS) is an important mechanism enabling the dynamic compartmentalization of macromolecules, including complex polymers such as proteins and nucleic acids, and occurs as a function of the physicochemical environment. In the model plant, Arabidopsis thaliana, LLPS by the protein EARLY FLOWERING3 (ELF3) occurs in a temperature-sensitive manner and controls thermoresponsive growth. ELF3 contains a largely unstructured prion-like domain (PrLD) that acts as a driver of LLPS in vivo and in vitro. The PrLD contains a poly-glutamine (polyQ) tract, whose length varies across natural Arabidopsis accessions. Here, we use a combination of biochemical, biophysical, and structural techniques to investigate the dilute and condensed phases of the ELF3 PrLD with varying polyQ lengths. We demonstrate that the dilute phase of the ELF3 PrLD forms a monodisperse higher-order oligomer that does not depend on the presence of the polyQ sequence. This species undergoes LLPS in a pH- and temperature-sensitive manner and the polyQ region of the protein tunes the initial stages of phase separation. The liquid phase rapidly undergoes aging and forms a hydrogel as shown by fluorescence and atomic force microscopies. Furthermore, we demonstrate that the hydrogel assumes a semiordered structure as determined by small-angle X-ray scattering, electron microscopy, and X-ray diffraction. These experiments demonstrate a rich structural landscape for a PrLD protein and provide a framework to describe the structural and biophysical properties of biomolecular condensates.
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低复杂性蛋白段的原子结构揭示了组装网络的扭结β薄片。
DOI：
10.1126/science.aan6398
发表时间：
2018-02-09
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Hughes MP;Sawaya MR;Boyer DR;Goldschmidt L;Rodriguez JA;Cascio D;Chong L;Gonen T;Eisenberg DS
通讯作者：
Eisenberg DS
影响因子：
3
作者：
Chu X;Sun T;Li Q;Xu Y;Zhang Z;Lai L;Pei J
通讯作者：
Pei J
DOI：
10.1016/j.jbc.2021.101194
发表时间：
2021-10
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hughes MP;Goldschmidt L;Eisenberg DS
通讯作者：
Eisenberg DS
影响因子：
3.3
作者：
Das, Suman;Amin, Alan N.;Chan, Hue Sun
通讯作者：
Chan, Hue Sun
影响因子：
2.9
作者：
Bellingham, CM;Lillie, MA;Keeley, FW
通讯作者：
Keeley, FW