Understanding Influenza

Methods in molecular biology, Год журнала: 2025, Номер unknown, С. 1 - 26

Опубликована: Янв. 1, 2025

Язык: Английский

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Role of the Psi Packaging Signal and Dimerization Initiation Sequence in the Organization of Rous Sarcoma Virus Gag-gRNA Co-Condensates

Viruses, Год журнала: 2025, Номер 17(1), С. 97 - 97

Опубликована: Янв. 13, 2025

Retroviral genome selection and virion assembly remain promising targets for novel therapeutic intervention. Recent studies have demonstrated that the Gag proteins of Rous sarcoma virus (RSV) human immunodeficiency type-1 (HIV-1) undergo nuclear trafficking, colocalize with nascent genomic viral RNA (gRNA) at transcription sites, may interact host factors, display biophysical properties characteristic biomolecular condensates. In present work, we utilized a controlled in vitro condensate assay advanced imaging approaches to investigate effects interactions between RSV condensates nonviral RNAs on abundance organization. We observed psi (Ψ) packaging signal dimerization initiation sequence (DIS) had stabilizing condensates, while lacking these features promoted or antagonized condensation, depending local protein concentration architecture. An containing Ψ, DIS, linkage structure (DLS) is capable stable dimer formation was act as bridge These observations suggest additional, condensate-related roles Gag-Ψ binding, gRNA dimerization, dimerization/multimerization packaging, representing significant step forward our understanding how collectively facilitate efficient packaging.

Язык: Английский

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Fluorescence Loss After Photoactivation (FLAPh): A Pulse-Chase Cellular Assay for Understanding Kinetics and Dynamics of Viral Inclusions

Methods in molecular biology, Год журнала: 2025, Номер unknown, С. 125 - 140

Опубликована: Янв. 1, 2025

Язык: Английский

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Controlled and orthogonal partitioning of large particles into biomolecular condensates

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Апрель 14, 2025

Язык: Английский

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Protein–RNA condensation kinetics via filamentous nanoclusters

Protein Science, Год журнала: 2025, Номер 34(6)

Опубликована: Май 24, 2025

Abstract Protein–RNA phase separation is at the center of membraneless biomolecular condensates governing cell physiology and pathology. Using an archetypical viral protein–RNA condensation model, we determined sequence events that starts with sub‐second formation a protomer two RNAs per protein dimer. Association additional RNA molecules to weaker secondary binding sites in this kickstarts crystallization‐like assembly molecular condensate. Primary nucleation faster than sum growth, which multistep process. nuclei grow over hundreds seconds into filaments subsequently nanoclusters approximately 600 nm diameter. Cryoelectron microscopy reveals internal structure formed by incoming layers made ribonucleoprotein oligomers, reminiscent genome packing nucleocapsid. These progress liquid condensate droplets undergo further partial coalescence yield typical hydrogel‐like coacervates may represent scaffold large factory infected cells. Our integrated experimental kinetic investigation exposes rate‐limiting steps structures along key biological pathway present across life kingdoms.

Язык: Английский

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Controlled and orthogonal partitioning of large particles into biomolecular condensates

Опубликована: Июль 16, 2024

Abstract Biomolecular condensates arising from liquid-liquid phase separation contribute to diverse cellular processes, such as gene expression. Partitioning of client molecules into is critical regulating the composition and function condensates. Previous studies suggest that size limits partitioning, with dextrans >5 nm excluded Here, we asked whether larger particles, macromolecular complexes, can partition based on particle-condensate interactions. We sought discover biophysical principles govern particle inclusion in or exclusion using polymer nanoparticles tailored surface chemistries models complexes. Particles coated polyethylene glycol (PEG) did not next leveraged PEGylated particles an inert platform which conjugated specific adhesive moieties. functionalized biotin partitioned containing streptavidin, driven by high-affinity biotin-streptavidin binding. Oligonucleotide-decorated exhibited varying degrees partitioning condensates, depending condensate composition. oligonucleotide-coated was tuned altering salt concentration, oligonucleotide length, density. Remarkably, beads distinct orthogonally immiscible Based our experiments, conclude arbitrarily large controllably biomolecular given sufficiently strong condensate-particle interactions, a conclusion also supported coarse-grained molecular dynamics simulations theory. These findings may provide insights how various processes are achieved clients well offer design for development drug delivery systems selectively target disease-related Significance Statement subcellular compartments recruit exclude molecules, even though lack enclosing membrane. Many biochemical reconstitution experiments have investigated mechanisms membraneless organelles control modeling cells spatiotemporally components regulate functions. One outstanding question strictly limited size. In this work, engineered sizes functionalities measured these variables determine partitioning. observed controlled orthogonal several types, Molecular recapitulated key results. Our work advances understanding regulated, nanoparticle toolbox inspire delivery.

Язык: Английский

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Liquid-liquid phase separation in viral infection: from the occurrence and function to treatment potentials

Colloids and Surfaces B Biointerfaces, Год журнала: 2024, Номер 246, С. 114385 - 114385

Опубликована: Ноя. 17, 2024

Язык: Английский

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Out of the Dark, into the Light: Metabolic Fluorescent Labeling of Nucleic Acids

ChemMedChem, Год журнала: 2024, Номер 19(15)

Опубликована: Май 7, 2024

This review outlines recent advances in live-cell imaging techniques for nucleic acids. We describe the evolution of these methods, particularly highlighting development metabolic labeling approaches compatible with living systems using fluorescence-based labeling.

Язык: Английский

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Protein-RNA condensation kinetics via filamentous nanoclusters

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Ноя. 8, 2024

Abstract Protein-RNA phase separation is at the center of membraneless biomolecular condensates governing cell physiology and pathology. Using an archetypical viral protein-RNA condensation model, we determined sequence events that starts with sub-second formation a protomer two RNAs per protein dimer. Association additional RNA molecules to weaker secondary binding sites in this kickstarts crystallization-like assembly molecular condensate. Primary nucleation faster than sum growth, which multistep process. nuclei grow over hundreds seconds into filaments subsequently nanoclusters circa 600 nm diameter. Cryoelectron microscopy reveals internal structure formed by incoming layers made ribonucleoprotein oligomers, reminiscent genome packing nucleocapsid. These progress liquid condensate droplets undergo further partial coalescence yield typical hydrogel-like coacervates may represent scaffold large factory infected cells. Our integrated experimental kinetic investigation exposes rate limiting steps structures along key biological pathway present across life kingdoms.

Язык: Английский

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Biological complexity of the phase-separated protein states

Elsevier eBooks, Год журнала: 2024, Номер unknown, С. 407 - 421

Опубликована: Ноя. 22, 2024

Язык: Английский

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