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Updated: Dec 3, 2025

Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
Published on: May 26, 2013
Function of a viral genome packaging motor from bacteriophage T4 is insensitive to DNA sequence
Youbin Mo1, Nicholas Keller1, Damian delToro1
1Department of Physics, University of California, San Diego, La Jolla, CA 92093, USA.
Abstract:
Many viruses employ ATP-powered motors during assembly to translocate DNA into procapsid shells. Previous reports raise the question if motor function is modulated by substrate DNA sequence: (i) the phage T4 motor exhibits large translocation rate fluctuations and pauses and slips; (ii) evidence suggests that the phage phi29 motor contacts DNA bases during translocation; and (iii) one theoretical model, the 'B-A scrunchworm', predicts that 'A-philic' sequences that transition more easily to A-form would alter motor function. Here, we use single-molecule optical tweezers measurements to compare translocation of phage, plasmid, and synthetic A-philic, GC rich sequences by the T4 motor. We observed no significant differences in motor velocities, even with A-philic sequences predicted to show higher translocation rate at high applied force. We also observed no significant changes in motor pausing and only modest changes in slipping. To more generally test for sequence dependence, we conducted correlation analyses across pairs of packaging events. No significant correlations in packaging rate, pausing or slipping versus sequence position were detected across repeated measurements with several different DNA sequences. These studies suggest that viral genome packaging is insensitive to DNA sequence and fluctuations in packaging motor velocity, pausing and slipping are primarily stochastic temporal events.
Insights
Viral DNA packaging motors show no sequence-specific behavior. Studies reveal that motor velocity, pausing, and slipping during DNA translocation are mainly random, not influenced by specific DNA sequences.
Area of Science:
- Virology
- Molecular Biology
- Biophysics
Background:
- Viruses utilize ATP-powered motors for DNA translocation into procapsid shells during assembly.
- The influence of DNA sequence on viral motor function remains an open question, with some studies suggesting potential sequence-specific interactions.
Purpose of the Study:
- To investigate whether the DNA sequence affects the function of the phage T4 viral DNA packaging motor.
- To determine if specific DNA sequences, such as A-philic or GC-rich sequences, alter motor translocation rates, pausing, or slipping.
Main Methods:
- Single-molecule optical tweezers were used to measure the translocation of various DNA sequences (phage, plasmid, synthetic A-philic, GC-rich) by the T4 motor.
- Correlation analyses were performed across multiple packaging events to assess sequence dependence.
Main Results:
- No significant differences in motor velocity were observed across different DNA sequences, including A-philic sequences.
- Motor pausing showed no significant changes, and slipping exhibited only modest alterations regardless of DNA sequence.
- Correlation analyses did not reveal significant sequence-position-dependent effects on packaging rate, pausing, or slipping.
Conclusions:
- Viral genome packaging by the T4 motor appears to be largely insensitive to the substrate DNA sequence.
- Observed fluctuations in motor velocity, pausing, and slipping are primarily stochastic temporal events rather than sequence-driven phenomena.
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