Single-Molecule Studies of Exonucleases: Following Cleavage Actions One Step at a Time

  • Gwangrog LeeEmail author
Part of the Biological and Medical Physics, Biomedical Engineering book series (BIOMEDICAL)


Nucleases catalyze numerous biological processes ranging from DNA replication, recombination, and repair, to RNA processing and degradation. These enzymes cleave the bonds between two adjacent polymer residues via hydrolysis, involving several catalytic steps. However, multi-step reactions are inherently difficult to examine using bulk assays due to ensemble averaging. One way to unravel the enzymatic steps involved in a process is to follow the path of catalytic reaction and enzymatic motion one molecule or one step at a time. Recent progress in single-molecule techniques has enabled very accurate measurements of inter and intramolecular motion on a nanometer scale. Single-molecule methods for studying nucleases include fluorescence imaging, optical tweezers, and flow-stretching techniques. Single-molecule dynamic tools have revolutionized the approach of studying biochemical reactions and have provided many new insights into enzyme kinetics and thermodynamics, reshaping the view of the underlying biochemical reactions. In this review, I emphasize the strength of single-molecule techniques, which are capable of dissecting the whole process into fine catalytic steps of the reaction. I further review novel mechanistic insights into how nucleases function.


RNA DNA Exonuclease Nuclease Degradation Polymerization 



This work was supported by the GIST Research Institute (GRI) and the National Research Foundation of Korea (NRF) grant funded by the Korea government (NRF-2016R1A2B4014556) and by the grant from the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (HA17C0031: #1720050).


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Life Sciences, Gwangju Institute of Science and TechnologyGwangjuKorea

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