Membraneless organelles and liquid-liquid phase separation – methods for their characterisation

Area of Science:

• Cell Biology
• Biophysics
• Biochemistry

Background:

• Membraneless organelles (MLOs) are vital intracellular compartments involved in numerous cellular processes, including stress response.
• MLOs form through liquid-liquid phase separation (LLPS), a biophysical phenomenon driven by thermodynamics.
• These structures are found in both the cytoplasm and membrane-bound organelles, notably the nucleus.

Purpose of the Study:

• To elucidate the biophysical principles governing the formation and function of MLOs.
• To provide an overview of experimental techniques applicable to MLO research.
• To highlight the role of intrinsically disordered regions (IDRs) and RNA in MLO assembly.

Main Methods:

• The study reviews existing literature on MLO formation and function.
• It discusses principles of liquid-liquid phase separation (LLPS).
• It covers various experimental techniques for studying MLOs, including biochemical and biological approaches.

Main Results:

• MLOs are formed by the spontaneous phase separation of proteins with intrinsically disordered regions (IDRs) and RNA.
• LLPS provides a mechanism for dynamic compartmentalization within cells.
• Understanding MLO formation is key to comprehending cellular physiology and stress responses.

Conclusions:

• MLOs are essential for cellular functions and are formed via LLPS.
• Intrinsically disordered proteins and RNA play critical roles in MLO assembly.
• A diverse range of experimental methods are available for studying these dynamic cellular structures.