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Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins
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Structural basis for DNA recognition by a viral genome-packaging machine.

Maria Chechik1,2, Sandra J Greive1,2, Alfred A Antson1,2

  • 1York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5DD, United Kingdom.

Proceedings of the National Academy of Sciences of the United States of America
|August 8, 2024
PubMed
Summary

Small terminase proteins recognize viral DNA through an unexpected mechanism. DNA passes through a central tunnel, with sequence recognition occurring as it exits, enabling precise genome packaging initiation.

Keywords:
DNA packagingbacteriophage HK97virus assembly

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Area of Science:

  • Virology
  • Structural Biology
  • Molecular Biology

Background:

  • DNA recognition by terminase proteins is essential for double-stranded DNA virus assembly and genome packaging.
  • The precise molecular mechanism of viral DNA recognition by small terminase proteins has remained largely unknown.

Purpose of the Study:

  • To elucidate the DNA recognition mechanism of the small terminase protein from bacteriophage HK97.
  • To identify the minimal DNA sequence recognized by the small terminase and determine the structure of the complex.

Main Methods:

  • Identification of the minimal DNA recognition site in the bacteriophage HK97 genome.
  • Cryo-electron microscopy (cryo-EM) to determine the structure of the small terminase-DNA complex.

Main Results:

  • The small terminase utilizes an unconventional mechanism where DNA transits through its central tunnel.
  • Sequence-specific recognition occurs as the DNA emerges from the tunnel, mediated by a transient substructure.
  • This interaction allows continuous engagement and sequence monitoring, enabling precise initiation and termination of genome packaging.

Conclusions:

  • The study reveals a novel mechanism for viral DNA recognition and packaging initiation by small terminase proteins.
  • This finding provides critical insights into the molecular basis of viral capsid assembly and DNA packaging in bacteriophages.