Clicking viruses—with chemistry toward mechanisms in infection

Journal of Virology, Год журнала: 2025, Номер unknown

Опубликована: Май 14, 2025

ABSTRACT Viruses subvert cells and evade host defense. They emerge unpredictably threaten humans livestock through their genetic phenotypic diversity. Despite more than 100 years since the discovery of viruses, molecular underpinnings virus infections are incompletely understood. The introduction new methodologies into field, such as that click chemistry some 10 ago, keeps uncovering facets viruses. Click uses bio-orthogonal reactions on chemical probes couples nucleic acids, proteins, lipids with tractable labels, fluorophores for single-cell single-molecule imaging, or biotin biochemical profiling infections. Its applications in single often achieve resolution provide important insights widely known phenomenon cell-to-cell infection variability. This review describes advances to unravel mechanisms a select set enveloped nonenveloped DNA RNA including adenovirus, herpesvirus, human immunodeficiency virus. It highlights recent breakthroughs viral DNA, RNA, protein, well host-derived lipid functions both live chemically fixed cells. discusses specific processes entry, uncoating, transcription, replication, packaging, assembly provides perspective explore cell biology, variability, genome organization particle.

Язык: Английский

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Structural and biochemical analysis of ligand binding in yeast Niemann–Pick type C1–related protein

Life Science Alliance, Год журнала: 2024, Номер 8(1), С. e202402990 - e202402990

Опубликована: Окт. 25, 2024

In eukaryotes, integration of sterols into the vacuolar/lysosomal membrane is critically dependent on Niemann-Pick type C (NPC) system. The system consists an integral protein, called NCR1 in yeast, and NPC2, a luminal soluble protein that transfers to N-terminal domain (NTD) before integration. Both proteins have been implicated sterol homeostasis yeast humans. Here, we investigate lipid binding NCR1/NPC2 transport determine crystal structures NTD. NTD binds both ergosterol cholesterol, with nearly identical conformations pocket. Apart from sterols, can also bind fluorescent analogs phosphatidylinositol, phosphatidylcholine, phosphatidylserine, as well sphingosine ceramide. We confirm multi-lipid scope using photo-crosslinkable clickable analogs, namely, pac-cholesterol, pac-sphingosine, pac-ceramide. Finally, reconstitute transfer pac-sphingosine NPC2 vitro. Collectively, our results support NPC work versatile machinery for vacuolar structurally diverse lipids, besides ergosterol.

Язык: Английский

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Structural and biochemical analysis of ligand binding in yeast Niemann-Pick type C1-related protein

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

Опубликована: Июнь 11, 2024

Abstract In eukaryotes, integration of sterols into the vacuolar/lysosomal membrane is critically dependent on Niemann-Pick type C (NPC) system. The system consists an integral protein, called NCR1 in yeast, and NPC2, a luminal soluble protein that transfers to N-terminal domain (NTD) before integration. Both proteins have been implicated sterol homeostasis yeast humans. Here, we investigate lipid binding NCR1/NPC2 transport determine crystal structures sterol-binding NTD. NTD binds both ergosterol cholesterol, with nearly identical conformations pocket. Apart from sterols, can also bind fluorescent analogs phosphatidylinositol, phosphatidylcholine phosphatidylserine as well sphingosine ceramide. We confirm multi-lipid scope using photo-crosslinkable clickable analogs, namely pac-cholesterol, pac-sphingosine pac-ceramide. Finally, reconstitute transfer NPC2 vitro . Collectively, our results support NPC work versatile machinery for vacuolar structurally diverse lipids, besides ergosterol. Summary blurb Results X-ray crystallography assays different lipids expand knowledge substrate C1-related yeast. GRAPHICAL ABSTRACT

Язык: Английский

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In Vitro Approaches for Detection of Protein-Lipid Interactions and Protein-to-Protein Transport of Lipids Using Bifunctional Lipid Analogs

Methods in molecular biology, Год журнала: 2024, Номер unknown, С. 249 - 258

Опубликована: Дек. 19, 2024

Язык: Английский

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