- Title Information
- Title
- Biophysical studies of phase separation: RNA-binding proteins at the nexus of neurodegeneration, cancer and viral infection
- Type of Resource (primo)
- dissertations
- Name:
Personal
- Name Part
- Perdikari, Theodora Myrto
- Role
- Role Term:
Text
- creator
- Name:
Personal
- Name Part
- Fawzi, Nicolas
- Role
- Role Term:
Text
- Advisor
- Name:
Personal
- Name Part
- Colvin, Vicki
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Tang, Jay
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Karniadakis, George
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Wheeler, Richard
- Role
- Role Term:
Text
- Reader
- Name:
Corporate
- Name Part
- Brown University. Biology and Medicine: Biomedical Engineering
- Role
- Role Term:
Text
- sponsor
- Origin Information
- Copyright Date
- 2022
- Physical Description
- Extent
- xix, 230 p.
- digitalOrigin
- born digital
- Note:
thesis
- Thesis (Ph. D.)--Brown University, 2022
- Genre (aat)
- theses
- Abstract
- Liquid-liquid phase separation (LLPS) is a prominent organizer of cellular biochemistry. Dozens of liquid-like condensates are found throughout the nucleus and the cytoplasm and the vast majority of them are enriched in RNA-binding proteins; the maestros of gene expression. Fused in Sarcoma (FUS) is an RNA-binding protein that phase separates via its low-complexity (LC) and arginine- glycine-glycine (RGG) domains. FUS localizes in liquid-like assemblies like stress granules and DNA- repair foci when the cell needs to combat stress and DNA damage respectively. However, mutations and/or abnormal levels of post-translational modifications (PTMs) disorient physiologically relevant interactions of FUS with RNA and other binding partners leading to a number of neurodegenerative diseases and cancers with unmet therapeutic needs. Combining computer simulations with solution- state nuclear magnetic resonance (NMR) has emerged as an unparalleled method to obtain atomic- level details of liquid-like structures. In this thesis, the main focus is towards a deeper understanding of the biophysical rules governing the interplay of FUS with PTMs, RNA and condensate-modifying molecules. In Chapter 2, I use coarse-grained (CG) modeling to show how measuring the effect of PTMs like phosphorylation and acetylation on the single-chain properties of various systems including FUS is predictive of their propensity to phase separate. In Chapter 3, I decipher the behavior of SARS-CoV-2 nucleocapsid protein which phase separates with RNA as I was equipped to face this challenge during the course of my doctoral studies in LLPS. In Chapter 4, I use NMR and biochemical assays to probe how condensate-modifying molecules alter FUS structure, phase behavior and interactions with RNA. In Chapter 5, I demonstrate that arginine-specific contacts are necessary for FUS RGG-RNA interactions and methylation alters heterotypic interactions of FUS RGG with RNA and the LC domain. Collectively, my work provides a detailed molecular view into the effect of PTMs, RNA and drug-like molecules on RNA-binding protein assembly. This knowledge can be harnessed to design in-vitro drug-design studies that screen the effect of drug molecules against FUS LLPS and aggregation.
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01040325")
- Topic
- Nuclear magnetic resonance
- Subject
- Topic
- liquid-liquid phase separation
- Subject
- Topic
- Fused in Sarcoma
- Subject
- Topic
- SARS-CoV-2
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01086260")
- Topic
- RNA-protein interactions
- Subject
- Topic
- Biomolecular condensates
- Subject
- Topic
- Molecular Dynamics Simulation
- Language
- Language Term (ISO639-2B)
- English
- Record Information
- Record Content Source (marcorg)
- RPB
- Record Creation Date
(encoding="iso8601")
- 20220706
- Identifier:
DOI
- 10.26300/7fnh-8877