Publications by authors named "Belen Solano"

To counteract hand, foot, and mouth disease-causing viruses such as enterovirus A71 and coxsackievirus A6, virus-like particles (VLPs) have emerged as a leading contender for the development of a multivalent vaccine. However, VLPs have shown rapid conversion from a highly immunogenic state to a less immunogenic state and low particle integrity lifetimes compared to inactivated virus vaccines, thus raising concerns about their overall stability. Here, we produce VLPs to investigate capsid stability using cryogenic electron microscopy (cryo-EM), mass spectrometry (MS), biochemical assays, and atomic force microscopy (AFM).

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Eukaryotes have one replicative helicase known as CMG, which centrally organizes and drives the replisome, and leads the way at the front of replication forks. Obtaining a deep mechanistic understanding of the dynamics of CMG is critical to elucidating how cells achieve the enormous task of efficiently and accurately replicating their entire genome once per cell cycle. Single-molecule techniques are uniquely suited to quantify the dynamics of CMG due to their unparalleled temporal and spatial resolution.

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Integrated single-molecule force-fluorescence spectroscopy setups allow for simultaneous fluorescence imaging and mechanical force manipulation and measurements on individual molecules, providing comprehensive dynamic and spatiotemporal information. Dual-beam optical tweezers (OT) combined with a confocal scanning microscope form a force-fluorescence spectroscopy apparatus broadly used to investigate various biological processes, in particular, protein:DNA interactions. Such experiments typically involve imaging of fluorescently labeled proteins bound to DNA and force spectroscopy measurements of trapped individual DNA molecules.

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The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that has been reconstituted from purified proteins in ensemble biochemical studies.

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Article Synopsis
  • Viral RNA-dependent RNA polymerases (RdRps) are crucial targets for broad-spectrum antiviral drugs, and a specific compound, T-1106 triphosphate, influences their function by causing delays in RNA synthesis.
  • Using magnetic tweezers, researchers found that the RdRp of enterovirus A-71 utilizes a "backtracked" state to facilitate processes like copy-back RNA synthesis and homologous recombination during viral replication.
  • The study indicates that pyrazine-carboxamide ribonucleotides enhance these processes without causing harmful mutations, suggesting they can promote the creation of defective viral genomes, presenting a new potential for antiviral therapies.
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DNA replication in eukaryotes initiates at many origins distributed across each chromosome. Origins are bound by the origin recognition complex (ORC), which, with Cdc6 and Cdt1, recruits and loads the Mcm2-7 (MCM) helicase as an inactive double hexamer during G1 phase. The replisome assembles at the activated helicase in S phase.

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Article Synopsis
  • Optical trapping of tiny particles is widely used in nanoscience, but traditional materials often limit the forces and precision achievable.
  • The study introduces single-crystal rutile TiO, which has unique optical properties and stability, allowing for better momentum transfer.
  • Rutile TiO nanocylinders can generate strong forces and torques with low laser power, exceeding the performance of other materials and enhancing control in nanoscale applications.
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Studying the dynamics of intracellular processes and investigating the interaction of individual macromolecules in live cells is one of the main objectives of cell biology. These macromolecules move, assemble, disassemble, and reorganize themselves in distinct manners under specific physiological conditions throughout the cell cycle. Therefore, in vivo experimental methods that enable the study of individual molecules inside cells at controlled culturing conditions have proved to be powerful tools to obtain insights into the molecular roles of these macromolecules and how their individual behavior influence cell physiology.

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Article Synopsis
  • Titanium dioxide (TiO2) is crucial for various optical and electronic applications, especially in its single-crystalline form, which outperforms its other phases.
  • A new top-down fabrication method enables the production of high-aspect-ratio single-crystal TiO2 nanostructures with specific side profiles, enhancing its usability.
  • This advancement includes a functionalization strategy that effectively coats these structures with biomolecules, enabling the creation of DNA-tethered TiO2 nanocylinders for innovative applications like optical torque tools.
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