Probing Centromere Plasticity and Chromatin Remodeling Using Single-Molecule Techniques

The centromere functions as an essential interactive hub for mediating the equal distribution of sister chromatids during eukaryotic cell division and is subject to cell cycle-specific plasticity regulation. To ensure the replication, transcription, and repair of genetic information, chromatin remodeling is an indispensable molecular biology event. Both centromere plasticity and chromatin remodeling involve numerous protein components (e.g., Constitutive Centromere-Associated Network (CCAN), SWItch/Sucrose Non-Fermentable (SWI/SNF)) and exhibit highly dynamic spatiotemporal regulation, posing significant challenges for traditional ensemble approaches in elucidating their detailed mechanisms. In recent years, single-molecule techniques have revolutionized the study of chromatin remodeling and kinetochore assembly. By capturing real-time, high-resolution dynamics of molecular interactions, these methods uncover mechanistic insights inaccessible to conventional approaches. This review summarizes key advances and emerging paradigms enabled by single-molecule analysis tools, highlighting how they are reshaping our understanding of the mechanism of mitotic fidelity and epigenetic regulation, and providing a roadmap for future research in these fields.