Formation of Biomolecular Condensates in Bacteria by Tuning Protein Electrostatics.
Formation of Biomolecular Condensates in Bacteria by Tuning Protein Electrostatics.
复制标题
DOI：
10.1021/acscentsci.0c01146
复制
发表时间：
2020-12-23
影响因子：
18.2
通讯作者：
Obermeyer AC
中科院分区：
文献类型：
--
作者：
Yeong V;Werth EG;Brown LM;Obermeyer AC
虽然真核细胞有无数的膜结合细胞器，能够分离不同的化学环境，但原核细胞缺乏这些明确的反应血管。生物分子凝聚体--没有膜的细胞器--提供了一种无物理屏障的细胞组织策略，同时允许细胞动态、灵敏的组织。众所周知，本质上无序的蛋白质结构域通过液-液相分离来驱动缩合物的形成；然而，球状蛋白质结构域在细胞内相分离中的作用仍然知之甚少。我们假设球状蛋白质的总电荷将决定凝聚体的形成和浓度，并系统地用大肠杆菌中的超负荷蛋白质和核酸来探索这一假说。在这项研究中，我们证明了凝聚体是通过工程蛋白质和RNA之间的静电相互作用形成的，并且这些凝聚体是动态的，只浓缩特定的核酸和蛋白质成分。在这里，我们提出了一个简单的基于蛋白质电荷的相分离模型，可以用来预测细胞内凝聚物的形成。有了这些指导方针，我们已经为设计具有可调节控制球状蛋白质功能的功能性合成无膜细胞器铺平了道路。工程超阳离子蛋白质在体外和细菌中与核酸进行液态−液相分离，形成选择性的生物分子凝聚体。
While eukaryotic cells have a myriad of membrane-bound organelles enabling the isolation of different chemical environments, prokaryotic cells lack these defined reaction vessels. Biomolecular condensates—organelles that lack a membrane—provide a strategy for cellular organization without a physical barrier while allowing for the dynamic, responsive organization of the cell. It is well established that intrinsically disordered protein domains drive condensate formation via liquid–liquid phase separation; however, the role of globular protein domains on intracellular phase separation remains poorly understood. We hypothesized that the overall charge of globular proteins would dictate the formation and concentration of condensates and systematically probed this hypothesis with supercharged proteins and nucleic acids in E. coli. Within this study, we demonstrated that condensates form via electrostatic interactions between engineered proteins and RNA and that these condensates are dynamic and only enrich specific nucleic acid and protein components. Herein, we propose a simple model for the phase separation based on protein charge that can be used to predict intracellular condensate formation. With these guidelines, we have paved the way to designer functional synthetic membraneless organelles with tunable control over globular protein function. Engineered supercationic proteins undergo liquid−liquid phase separation with nucleic acids in vitro and in bacteria to form selective biomolecular condensates.
登录
查看更多内容
影响因子：
12.3
作者：
Dalgleish, Raymond;Flicek, Paul;Maglott, Donna R.
通讯作者：
Maglott, Donna R.
DOI：
10.1073/pnas.1509317112
发表时间：
2015-09-22
影响因子：
11.1
作者：
Berry, Joel;Weber, Stephanie C.;Brangwynne, Clifford P.
通讯作者：
Brangwynne, Clifford P.
影响因子：
64.8
作者：
Larson AG;Elnatan D;Keenen MM;Trnka MJ;Johnston JB;Burlingame AL;Agard DA;Redding S;Narlikar GJ
通讯作者：
Narlikar GJ
影响因子：
19
作者：
Boeynaems S;Alberti S;Fawzi NL;Mittag T;Polymenidou M;Rousseau F;Schymkowitz J;Shorter J;Wolozin B;Van Den Bosch L;Tompa P;Fuxreiter M
通讯作者：
Fuxreiter M
影响因子：
14.8
作者：
Lee, Matthew J.;Mantell, Judith;Warren, Martin J.
通讯作者：
Warren, Martin J.