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Non-equilibrium structural dynamics of supercoiled DNA plasmids exhibits asymmetrical relaxation.


ABSTRACT: Many cellular processes occur out of equilibrium. This includes site-specific unwinding in supercoiled DNA, which may play an important role in gene regulation. Here, we use the Convex Lens-induced Confinement (CLiC) single-molecule microscopy platform to study these processes with high-throughput and without artificial constraints on molecular structures or interactions. We use two model DNA plasmid systems, pFLIP-FUSE and pUC19, to study the dynamics of supercoiling-induced secondary structural transitions after perturbations away from equilibrium. We find that structural transitions can be slow, leading to long-lived structural states whose kinetics depend on the duration and direction of perturbation. Our findings highlight the importance of out-of-equilibrium studies when characterizing the complex structural dynamics of DNA and understanding the mechanisms of gene regulation.

SUBMITTER: Shaheen C 

PROVIDER: S-EPMC8934633 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Non-equilibrium structural dynamics of supercoiled DNA plasmids exhibits asymmetrical relaxation.

Shaheen Cynthia C   Hastie Cameron C   Metera Kimberly K   Scott Shane S   Zhang Zhi Z   Chen Sitong S   Gu Gracia G   Weber Lisa L   Munsky Brian B   Kouzine Fedor F   Levens David D   Benham Craig C   Leslie Sabrina S  

Nucleic acids research 20220301 5


Many cellular processes occur out of equilibrium. This includes site-specific unwinding in supercoiled DNA, which may play an important role in gene regulation. Here, we use the Convex Lens-induced Confinement (CLiC) single-molecule microscopy platform to study these processes with high-throughput and without artificial constraints on molecular structures or interactions. We use two model DNA plasmid systems, pFLIP-FUSE and pUC19, to study the dynamics of supercoiling-induced secondary structura  ...[more]

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