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Updated: May 21, 2026

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

A single-molecule Hershey-Chase experiment.

David Van Valen1, David Wu, Yi-Ju Chen

  • 1Division of Engineering and Applied Sciences, California Institute of Technology, Pasadena, CA 91125, USA.

Current Biology : CB
|June 26, 2012
PubMed
Summary
This summary is machine-generated.

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Researchers visualized bacteriophage DNA ejection into E. coli cells, finding in vivo ejection takes minutes, unlike rapid in vitro DNA release. This suggests cell-internal factors control phage DNA translocation during infection.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biophysics

Background:

  • The mechanism of DNA translocation from bacteriophages into host cells remains largely unknown.
  • Previous studies relied on bulk measurements or in vitro single-molecule assays for DNA ejection.
  • Understanding phage DNA injection is crucial for deciphering early infection stages and polymer translocation.

Purpose of the Study:

  • To directly visualize and quantify single bacteriophage DNA ejection in vivo during infection of Escherichia coli.
  • To compare the dynamics of in vivo DNA ejection with in vitro single-molecule ejection.
  • To elucidate the factors controlling bacteriophage DNA translocation.

Main Methods:

  • Direct visualization of single bacteriophages infecting individual Escherichia coli cells using advanced microscopy.

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Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG
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Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG

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Last Updated: May 21, 2026

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

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Published on: August 27, 2014

Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1
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Published on: July 26, 2024

  • Quantitative analysis of DNA ejection time, velocity, and variability in real-time.
  • Comparison of in vivo ejection dynamics with in vitro single-molecule DNA ejection experiments.

    • Main Results:

    • Bacteriophage lambda (λ) exhibits a mean in vivo DNA ejection time of approximately 5 minutes, with considerable cell-to-cell variability and pausing events.
    • In vitro single-molecule ejections are significantly faster and more uniform, completing within 10 seconds.
    • Ejection velocity in vivo correlates with the amount of DNA ejected, suggesting internal cellular control, whereas in vitro ejection depends on DNA remaining in the capsid.

    Conclusions:

    • In vivo bacteriophage DNA ejection is a slower, more variable process than in vitro ejection, indicating the influence of host cell factors.
    • The observed correlation between ejection velocity and DNA amount ejected in vivo challenges purely repulsion-based models and supports a model dominated by cell-internal processes.
    • This study provides critical insights into the early stages of phage infection and the complex problem of polymer translocation across cellular barriers.