Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes.

ABSTRACT: The ability of proteins involved in eukaryotic DNA replication to overcome obstacles - such as protein and DNA 'roadblocks' - is critical for ensuring faithful genome duplication. G-quadruplexes are higher-order nucleic acid structures that form in guanine-rich regions of DNA and have been shown to act as obstacles, interfering with genomic maintenance pathways. This study introduces a real-time, fluorescence microscopy-based method to observe DNA polymerase interactions with G-quadruplex structures. Short, primed DNA oligonucleotides containing a G-quadruplex were immobilized on functionalized glass coverslips within a microfluidic flow cell. Fluorescently labeled DNA polymerases were introduced, allowing their behavior and stoichiometry to be monitored over time. This approach enabled th

SUBMITTER: Kusi-Appauh N 

PROVIDER: S-EPMC12169813 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature