A5074141541- RNA Research and Splicing
- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- Amyotrophic Lateral Sclerosis Research
- Nuclear Structure and Function
- Neurogenetic and Muscular Disorders Research
- Viral Infections and Immunology Research
- RNA regulation and disease
- Metabolism, Diabetes, and Cancer
- Pancreatic function and diabetes
- Ancient Mediterranean Archaeology and History
- Modular Robots and Swarm Intelligence
- Classical Antiquity Studies
- RNA Interference and Gene Delivery
- biodegradable polymer synthesis and properties
- PARP inhibition in cancer therapy
- Cancer, Hypoxia, and Metabolism
University of California, San Diego
2022-2024
Johns Hopkins University
2019-2023
Portland State University
2013
Biomolecular condensates often consist of intrinsically disordered protein and RNA molecules, which together promote the formation membraneless organelles in cells. The nucleation, condensation, maturation is a critical yet poorly understood process. Here, we present single-molecule accompanying ensemble methods to quantify these processes more comprehensively. In particular, focus on how properly design execute nucleation assay, detect signals arising from individual units fluorescently...
Messenger RNAs (mRNAs) interact with RNA-binding proteins (RBPs) in diverse ribonucleoprotein complexes (RNPs) during distinct life-cycle stages for their processing and maturation. While substantial attention has focused on understanding RNA regulation by assigning proteins, particularly RBPs, to specific substrates, there been considerably less exploration leveraging protein-protein interaction (PPI) methodologies identify study the role of mRNA stages. To address this gap, we generated an...
Abstract RNA-binding proteins (RBPs) have pivotal functions in RNA metabolism, but current methods are limited retrieving RBP-RNA interactions within endogenous biological contexts. Here, we develop INSCRIBE ( IN situ S ensitive C apture of R NA-protein I nteractions B iological E nvironments), circumventing the challenges through labeling by precisely directing a purified APOBEC1-nanobody fusion to RBP interest. This method enables highly specific site identification across diverse range...
Abstract The glycolytic enzyme fructose 1,6‐(bis)phosphate aldolase (aldolase) is not only required for efficient utilization of glucose and fructose, but also cytoskeletal functions like cytokinesis cell motility. These differing roles are mediated by distinct discrete binding interactions with aldolase's many partners, including actin filaments, Wiskott‐Aldrich Syndrome protein (WASP), Sorting Nexin 9 (SNX9). How these coordinated on the homotetramer 160 kDa unclear. In this study,...
Liquid-liquid phase separation of intrinsically disordered proteins is known to underlie diverse pathologies such as neurodegeneration, cancer, and aging. The nucleation step condensate formation critical importance for understanding how healthy disease-associated condensates differ. Here, we describe four orthogonal single-molecule techniques that enable molecular tracking the RNA-protein interaction, RNA-induced oligomerization, kinetics nucleation. These approaches allow researchers...
Poly(ADP-ribose) (PAR) is an RNA-like polymer that regulates increasing number of biological processes. Dysregulation PAR implicated in neurodegenerative diseases characterized by abnormal protein aggregation, including Amyotrophic Lateral Sclerosis (ALS). forms condensates with FUS, RNA-binding linked ALS, through unknown mechanism. Here, we demonstrate a strikingly low concentration (1 nM) sufficient to trigger condensation FUS near its physiological µM), which three orders magnitude lower...
The formation of pathogenic inclusions RNA-binding proteins in neurons is a hallmark amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD). One prominent protein these Fused sarcoma (FUS), over 70 mutations Fus are linked to ALS/FTLD. In patients, all heterozygous, indicating that the mutant drives pathology despite presence wild-type FUS. Here, we demonstrate ALS FUS glycine (G) strikingly drive droplets immiscible with whereas arginine (R) mutants form miscible...
Karyopherin-β2 (Kapβ2) is a nuclear import receptor (NIR) that localizes proteins bearing proline-tyrosine localization signal (PY-NLS) to the nucleus, including RNA-binding protein (RBP) FUS. Recent work has demonstrated NIRs can chaperone RBPs in vitro and cells, preventing reversing their self-assembly aggregation. However, when PY-NLS mutated, Kapβ2 no longer efficiently chaperones its cargo. Here, we focus on case of FUSP525L , an RBP NLS mutant associated with highly aggressive form...
Abstract Karyopherin-β2 (Kapβ2) is a nuclear-import receptor that recognizes proline-tyrosine nuclear localization signals (PY-NLSs) of diverse cytoplasmic cargo for transport to the nucleus. Kapβ2 include several disease-linked RNA-binding proteins (RBPs) with prion-like domains (PrLDs), such as FUS, TAF15, EWSR1, hnRNPA1, and hnRNPA2. These RBPs PrLDs are linked via pathology genetics debilitating degenerative disorders, including amyotrophic lateral sclerosis (ALS), frontotemporal...