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Single-Molecular Dissection of Liquid-Liquid Phase Transitions
Pravin Pokhrel1, Sagun Jonchhe1, Wei Pan1
1Department of Chemistry and Biochemistry, Kent State University, Kent, Ohio 44242, United States.
Journal of the American Chemical Society
|July 26, 2023
Summary
Poly-l-lysine (PLL) peptide interactions with nucleic acids drive liquid-liquid phase separation (LLPS) in cells. Nucleic acid rigidity influences LLPS condensate formation, offering insights into cellular organization.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophysics
Background:
- Peptide-nucleic acid interactions are fundamental to cellular processes like replication and transcription.
- Liquid-liquid phase separation (LLPS) forms membraneless organelles crucial for cellular functions.
- Understanding LLPS mechanisms is key to deciphering cellular organization and dysfunction.
Purpose of the Study:
- To investigate the molecular mechanisms of poly-l-lysine (PLL) and nucleic acid interactions during early-stage LLPS.
- To elucidate the role of nucleic acid properties in modulating LLPS condensate formation.
- To provide insights into the transition dynamics of LLPS condensates.
Main Methods:
- Single-molecule optical tweezers to monitor mechanical tension.
- Controlled addition of poly-l-lysine (PLL) to nucleic acid templates.
- Systematic variation of nucleic acid types (ssDNA, ssRNA, dsDNA) and sequences.
Main Results:
- Revealed a multistage LLPS process driven by long-range polyelectrolyte interactions.
- Demonstrated that nucleic acid rigidity negatively correlates with LLPS condensate formation.
- Constructed transition diagrams for PLL-nucleic acid condensates.
Conclusions:
- Nucleic acid rigidity is a critical factor controlling PLL-mediated LLPS.
- Findings offer a deeper understanding of LLPS condensate transitions.
- Potential for developing strategies to modulate LLPS-driven cellular functions.
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