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Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

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Title
Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique
Author(s)
Kang, Yujin; Bae, Subin; An, Soyeong; Ja Yil Lee
Publication Date
2022-03
Journal
Journal of Visualized Experiments, v.2022, no.181
Publisher
Journal of Visualized Experiments
Abstract
© 2022 JoVE Journal of Visualized Experiments.Chromatin is a higher-order structure that packages eukaryotic DNA. Chromatin undergoes dynamic alterations according to the cell cycle phase and in response to environmental stimuli. These changes are essential for genomic integrity, epigenetic regulation, and DNA metabolic reactions such as replication, transcription, and repair. Chromatin assembly is crucial for chromatin dynamics and is catalyzed by histone chaperones. Despite extensive studies, the mechanisms by which histone chaperones enable chromatin assembly remains elusive. Moreover, the global features of nucleosomes organized by histone chaperones are poorly understood. To address these problems, this work describes a unique single-molecule imaging technique named DNA curtain, which facilitates the investigation of the molecular details of nucleosome assembly by histone chaperones. DNA curtain is a hybrid technique that combines lipid fluidity, microfluidics, and total internal reflection fluorescence microscopy (TIRFM) to provide a universal platform for real-time imaging of diverse protein-DNA interactions.Using DNA curtain, the histone chaperone function of Abo1, the Schizosaccharomyces pombe bromodomain-containing AAA+ ATPase, is investigated, and the molecular mechanism underlying histone assembly of Abo1 is revealed. DNA curtain provides a unique approach for studying chromatin dynamics.
URI
https://pr.ibs.re.kr/handle/8788114/12952
DOI
10.3791/63501
ISSN
1940-087X
Appears in Collections:
Center for Genomic Integrity(유전체 항상성 연구단) > 1. Journal Papers (저널논문)
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