Single-Molecule Analysis and Engineering of DNA Motors

Sonisilpa Mohapatra, Chang Ting Lin, Xinyu A. Feng, Aakash Basu, Taekjip Ha

Research output: Contribution to journalReview article

Abstract

Molecular motors are diverse enzymes that transduce chemical energy into mechanical work and, in doing so, perform critical cellular functions such as DNA replication and transcription, DNA supercoiling, intracellular transport, and ATP synthesis. Single-molecule techniques have been extensively used to identify structural intermediates in the reaction cycles of molecular motors and to understand how substeps in energy consumption drive transitions between the intermediates. Here, we review a broad spectrum of single-molecule tools and techniques such as optical and magnetic tweezers, atomic force microscopy (AFM), single-molecule fluorescence resonance energy transfer (smFRET), nanopore tweezers, and hybrid techniques that increase the number of observables. These methods enable the manipulation of individual biomolecules via the application of forces and torques and the observation of dynamic conformational changes in single motor complexes. We also review how these techniques have been applied to study various motors such as helicases, DNA and RNA polymerases, topoisomerases, nucleosome remodelers, and motors involved in the condensation, segregation, and digestion of DNA. In-depth analysis of mechanochemical coupling in molecular motors has made the development of artificially engineered motors possible. We review techniques such as mutagenesis, chemical modifications, and optogenetics that have been used to re-engineer existing molecular motors to have, for instance, altered speed, processivity, or functionality. We also discuss how single-molecule analysis of engineered motors allows us to challenge our fundamental understanding of how molecular motors transduce energy.

Original languageEnglish (US)
JournalChemical Reviews
DOIs
StatePublished - Jan 1 2019

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Molecules
DNA
Mutagenesis
Nanopores
Nucleosomes
Chemical modification
Biomolecules
DNA-Directed DNA Polymerase
DNA-Directed RNA Polymerases
Transcription
Condensation
Atomic force microscopy
Energy utilization
Torque
Adenosine Triphosphate
Engineers
Enzymes

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Single-Molecule Analysis and Engineering of DNA Motors. / Mohapatra, Sonisilpa; Lin, Chang Ting; Feng, Xinyu A.; Basu, Aakash; Ha, Taekjip.

In: Chemical Reviews, 01.01.2019.

Research output: Contribution to journalReview article

Mohapatra, Sonisilpa ; Lin, Chang Ting ; Feng, Xinyu A. ; Basu, Aakash ; Ha, Taekjip. / Single-Molecule Analysis and Engineering of DNA Motors. In: Chemical Reviews. 2019.
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