Formation and functionalization of membraneless compartments in Escherichia coli

• Published in: Nature chemical biology Vol. 16; no. 10; pp. 1143 - 1148
• Main Authors: Wei, Shao-Peng, Qian, Zhi-Gang, Hu, Chun-Fei, Pan, Fang, Chen, Meng-Ting, Lee, Sang Yup, Xia, Xiao-Xia
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-10-2020; Nature Publishing Group
• Subjects: 631/92/469; 631/92/470; 631/92/552; Biochemical Engineering; Biochemistry; Biological activity; Bioorganic Chemistry; Cell Biology; Cell Membrane; Chemical reactions; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Compartments; Condensates; Cytosol - chemistry; Cytosol - metabolism; Dynamic Light Scattering; E coli; Escherichia coli; Escherichia coli - physiology; Eukaryotes; Fibroins - chemistry; Gene Expression Regulation, Bacterial; Green Fluorescent Proteins - chemistry; Liquid phases; Microscopy, Confocal; Microscopy, Electron, Transmission; Microscopy, Fluorescence; Nucleic acids; Organelles; Phase separation; Prokaryotes; Proteins; Silk; Structural proteins
• ISSN: 1552-4450; 1552-4469
• DOI: 10.1038/s41589-020-0579-9

Membraneless organelles formed by liquid–liquid phase separation of proteins or nucleic acids are involved in diverse biological processes in eukaryotes. However, such cellular compartments have yet to be discovered or created synthetically in prokaryotes. Here, we report the formation of liquid protein condensates inside the cells of prokaryotic
Escherichia coli
upon heterologous overexpression of intrinsically disordered proteins such as spider silk and resilin. In vitro reconstitution under conditions that mimic intracellular physiologically crowding environments of
E. coli
revealed that the condensates are formed via liquid–liquid phase separation. We also show functionalization of these condensates via targeted colocalization of cargo proteins to create functional membraneless compartments able to fluoresce and to catalyze biochemical reactions. The ability to form and functionalize membraneless compartments may serve as a versatile tool to develop artificial organelles with on-demand functions in prokaryotes for applications in synthetic biology.
Utilization of structural proteins with intrinsically disordered regions enables the formation of membraneless organelles in
Escherichia coli
through liquid–liquid phase separation, as well as their functionalization with active enzymes.