Engineering Circular Gliding of Actin Filaments Along Myosin-Patterned DNA Nanotube Rings to Study Long-Term Actin-Myosin Behaviors

Rizal Hariadi, Abhinav J. Appukutty, Sivaraj Sivaramakrishnan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Nature has evolved molecular motors that are critical in cellular processes occurring over broad time scales, ranging from seconds to years. Despite the importance of the long-term behavior of molecular machines, topics such as enzymatic lifetime are underexplored due to the lack of a suitable approach for monitoring motor activity over long time periods. Here, we developed an "O"-shaped Myosin Empowered Gliding Assay (OMEGA) that utilizes engineered micron-scale DNA nanotube rings with precise arrangements of myosin VI to trap gliding actin filaments. This circular gliding assay platform allows the same individual actin filament to glide over the same myosin ensemble (50-1000 motors per ring) multiple times. First, we systematically characterized the formation of DNA nanotubes rings with 4, 6, 8, and 10 helix circumferences. Individual actin filaments glide along the nanotube rings with high processivity for up to 12.8 revolutions or 11 min in run time. We then show actin gliding speed is robust to variation in motor number and independent of ring curvature within our sample space (ring diameter of 0.5-4 μm). As a model application of OMEGA, we then analyze motor-based mechanical influence on "stop-and-go" gliding behavior of actin filaments, revealing that the stop-to-go transition probability is dependent on motor flexibility. Our circular gliding assay may provide a closed-loop platform for monitoring long-term behavior of broad classes of molecular motors and enable characterization of motor robustness and long time scale nanomechanical processes.

Original languageEnglish (US)
Pages (from-to)8281-8288
Number of pages8
JournalACS Nano
Volume10
Issue number9
DOIs
StatePublished - Sep 27 2016
Externally publishedYes

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myosins
gliding
Myosins
Nanotubes
Actins
nanotubes
filaments
DNA
deoxyribonucleic acid
engineering
rings
Assays
platforms
Monitoring
circumferences
transition probabilities
helices
flexibility
curvature
traps

Keywords

  • actin gliding
  • DNA nanotechnology
  • molecular motors

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)

Cite this

Engineering Circular Gliding of Actin Filaments Along Myosin-Patterned DNA Nanotube Rings to Study Long-Term Actin-Myosin Behaviors. / Hariadi, Rizal; Appukutty, Abhinav J.; Sivaramakrishnan, Sivaraj.

In: ACS Nano, Vol. 10, No. 9, 27.09.2016, p. 8281-8288.

Research output: Contribution to journalArticle

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abstract = "Nature has evolved molecular motors that are critical in cellular processes occurring over broad time scales, ranging from seconds to years. Despite the importance of the long-term behavior of molecular machines, topics such as enzymatic lifetime are underexplored due to the lack of a suitable approach for monitoring motor activity over long time periods. Here, we developed an {"}O{"}-shaped Myosin Empowered Gliding Assay (OMEGA) that utilizes engineered micron-scale DNA nanotube rings with precise arrangements of myosin VI to trap gliding actin filaments. This circular gliding assay platform allows the same individual actin filament to glide over the same myosin ensemble (50-1000 motors per ring) multiple times. First, we systematically characterized the formation of DNA nanotubes rings with 4, 6, 8, and 10 helix circumferences. Individual actin filaments glide along the nanotube rings with high processivity for up to 12.8 revolutions or 11 min in run time. We then show actin gliding speed is robust to variation in motor number and independent of ring curvature within our sample space (ring diameter of 0.5-4 μm). As a model application of OMEGA, we then analyze motor-based mechanical influence on {"}stop-and-go{"} gliding behavior of actin filaments, revealing that the stop-to-go transition probability is dependent on motor flexibility. Our circular gliding assay may provide a closed-loop platform for monitoring long-term behavior of broad classes of molecular motors and enable characterization of motor robustness and long time scale nanomechanical processes.",
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