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20 August 2020 Restricted mobility of DNA packaged in phage phi29 viral proheads assessed by single-molecule optical tweezers measurements of DNA exit
Mounir Fizari, Nicholas Keller, Douglas E. Smith
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Proceedings Volume 11463, Optical Trapping and Optical Micromanipulation XVII; 1146303 (2020) https://doi.org/10.1117/12.2570185
Event: SPIE Nanoscience + Engineering, 2020, Online Only
Abstract
Many viruses contain DNA packed to such a high density that the mobility of the DNA inside the viral capsid is severely restricted, affecting the processes of DNA packaging and ejection. We study phage phi29, which uses an ATP-powered molecular motor to package DNA, by using optical tweezers to measure DNA ejection through the phi29 portal-motor channel after the removal of ATP during DNA packaging. We find that when initiated at low capsid filling levels, DNA exits faster than 10 kbp/s. When initiated at high filling levels, exit occurs with a dramatically reduced average velocity that decreases with increasing initial prohead filling. In individual exit measurements, complex dynamics and transient pausing are seen, which we attribute to the nonequilibrium DNA conformations thought to arise during DNA packaging. We also show that high concentration of Mg2+ slows exit dynamics, suggesting that the internal pressure of the confined DNA is the driving force for the ejection process.
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Mounir Fizari, Nicholas Keller, and Douglas E. Smith "Restricted mobility of DNA packaged in phage phi29 viral proheads assessed by single-molecule optical tweezers measurements of DNA exit", Proc. SPIE 11463, Optical Trapping and Optical Micromanipulation XVII, 1146303 (20 August 2020); https://doi.org/10.1117/12.2570185
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KEYWORDS
Optical testing
Optical tweezers
Packaging
Acquisition tracking and pointing
Luminescence
Viruses
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