Periodic Reporting for period 4 - LLPS-NMR (Nuclear magnetic resonance spectroscopy of liquid-liquid phase separation)
Reporting period: 2023-03-01 to 2024-08-31
The ERC project LLPS-NMR aimed to tackle a fundamental challenge in modern biology and chemistry: understanding the molecular mechanisms of liquid-liquid phase separation (LLPS) in biomolecules. LLPS is a thermodynamic phenomenon where molecules demix into condensed liquid-like phases, forming biomolecular condensates. These condensates play crucial roles in cellular organization, compartmentalization, and regulation of biochemical reactions, but their misregulation is implicated in diseases like Alzheimer's disease (AD), Parkinson’s disease (PD), and ALS. Despite their importance, LLPS processes were largely studied using low-resolution methods like light microscopy, leaving critical questions unanswered about the structure, dynamics, and regulation of proteins in these states.
Understanding LLPS is critical for:
-Disease Mechanisms: Misregulation of LLPS is linked to pathological aggregation in neurodegenerative diseases such as AD and PD. Insights into LLPS could lead to new diagnostic tools and treatments.
-Therapeutics: By uncovering how phase-separated states form and function, the project provides a foundation for designing therapeutic interventions targeting LLPS-related processes.
-Fundamental Biology: LLPS plays a role in essential cellular functions like gene expression, RNA metabolism, and stress responses. Gaining high-resolution insights transforms our understanding of how cells are spatially and temporally organized.
The overall objectives of the LLPS-NMR project were to:
-Develop novel NMR spectroscopy techniques to overcome the resolution barrier, advancing from micrometer to sub-nanometer scales for the study of phase-separated biomolecules.
-Elucidate the molecular structure and dynamics of intrinsically disordered proteins (IDPs) in liquid-like condensates.
-Disentangle the chemical reactions and interactions occurring within phase-separated compartments.
-Understand the regulatory mechanisms of LLPS, focusing on how post-translational modifications, nucleic acids, and other cellular factors influence phase behavior.
-Identify pathological mechanisms of LLPS dysregulation in diseases like Alzheimer’s disease.
Understanding LLPS is critical for:
-Disease Mechanisms: Misregulation of LLPS is linked to pathological aggregation in neurodegenerative diseases such as AD and PD. Insights into LLPS could lead to new diagnostic tools and treatments.
-Therapeutics: By uncovering how phase-separated states form and function, the project provides a foundation for designing therapeutic interventions targeting LLPS-related processes.
-Fundamental Biology: LLPS plays a role in essential cellular functions like gene expression, RNA metabolism, and stress responses. Gaining high-resolution insights transforms our understanding of how cells are spatially and temporally organized.
The overall objectives of the LLPS-NMR project were to:
-Develop novel NMR spectroscopy techniques to overcome the resolution barrier, advancing from micrometer to sub-nanometer scales for the study of phase-separated biomolecules.
-Elucidate the molecular structure and dynamics of intrinsically disordered proteins (IDPs) in liquid-like condensates.
-Disentangle the chemical reactions and interactions occurring within phase-separated compartments.
-Understand the regulatory mechanisms of LLPS, focusing on how post-translational modifications, nucleic acids, and other cellular factors influence phase behavior.
-Identify pathological mechanisms of LLPS dysregulation in diseases like Alzheimer’s disease.
The project achieved several breakthroughs:
-High-Resolution NMR for Phase-Separated States: Novel spatially-resolved NMR techniques were developed, enabling quantitative, label-free analysis of the internal structure and composition of biomolecular condensates.
-Insights into Tau and Alzheimer’s Disease: The project demonstrated how tau undergoes phase separation and transitions from liquid-like condensates into solid aggregates, revealing critical links between LLPS and pathological aggregation in Alzheimer’s disease.
-RNA Polymerase II and Transcriptional Regulation: The research uncovered how the C-terminal domain (CTD) of RNA polymerase II forms transcriptional hubs through LLPS, advancing understanding of gene expression regulation and its potential dysregulation in diseases.
-Regulatory Mechanisms of LLPS: The project identified how factors such as multivalent interactions, post-translational modifications, and RNA influence phase behavior, providing insights into the fine-tuning of condensate dynamics.
-Disease Implications and Therapeutic Potential: By linking LLPS to diseases like Alzheimer’s and Parkinson’s, the project laid the groundwork for targeting LLPS pathways therapeutically.
-High-Resolution NMR for Phase-Separated States: Novel spatially-resolved NMR techniques were developed, enabling quantitative, label-free analysis of the internal structure and composition of biomolecular condensates.
-Insights into Tau and Alzheimer’s Disease: The project demonstrated how tau undergoes phase separation and transitions from liquid-like condensates into solid aggregates, revealing critical links between LLPS and pathological aggregation in Alzheimer’s disease.
-RNA Polymerase II and Transcriptional Regulation: The research uncovered how the C-terminal domain (CTD) of RNA polymerase II forms transcriptional hubs through LLPS, advancing understanding of gene expression regulation and its potential dysregulation in diseases.
-Regulatory Mechanisms of LLPS: The project identified how factors such as multivalent interactions, post-translational modifications, and RNA influence phase behavior, providing insights into the fine-tuning of condensate dynamics.
-Disease Implications and Therapeutic Potential: By linking LLPS to diseases like Alzheimer’s and Parkinson’s, the project laid the groundwork for targeting LLPS pathways therapeutically.
The ERC LLPS-NMR project successfully bridged the gap between low-resolution studies of LLPS and the need for atomic-scale insights. By developing innovative NMR methodologies, it provided a transformative understanding of biomolecular condensates' structure, dynamics, and function. These findings have profound implications for biology, biomedicine, and therapeutic development, offering new avenues for addressing neurodegenerative diseases and other disorders associated with LLPS dysregulation.