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PreprintCited by 32Bacteriophages and microbial interactionsEnzyme Structure and FunctionAdvanced Electron Microscopy Techniques and Applications

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Insights into a viral motor: the structure of the HK97 packaging termination assembly

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Hawkins, Dorothy E.D.P., Bayfield, Oliver, Fung, Herman K.H., Grba, Daniel N, Huet, Alexis, Conway, James F., Antson, Alfred A.·2023·Nucleic Acids Research
DOI 10.1101/2023.02.24.529869PMC PMC10359639

Preprint — published as 10.1093/nar/gkad480

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Abstract

Double-stranded DNA viruses utilise machinery, made of terminase proteins, to package viral DNA into the capsid. For cos bacteriophage, a defined signal, recognised by small terminase, flanks each genome unit. Here we present the first structural data for a cos virus DNA packaging motor, assembled from the bacteriophage HK97 terminase proteins, procapsids encompassing the portal protein, and DNA containing a cos site. The cryo-EM structure is consistent with the packaging termination state adopted after DNA cleavage, with DNA density within the large terminase assembly ending abruptly at the portal protein entrance. Retention of the large terminase complex after cleavage of the short DNA substrate suggests that motor dissociation from the capsid requires headful pressure, in common with pac viruses. Interestingly, the clip domain of the 12-subunit portal protein does not adhere to C 12 symmetry, indicating asymmetry induced by binding of the large terminase/DNA. The motor assembly is also highly asymmetric, showing a ring of 5 large terminase monomers, tilted against the portal. Variable degrees of extension between N- and C-terminal domains of individual subunits suggest a mechanism of DNA translocation driven by inter-domain contraction and relaxation.

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Metadata

Authors
Hawkins, D. E. D. P., Bayfield, O., Fung, H. K. H., Grba, D. N., Huet, A., Conway, J. F., & Antson, A. A.
Affiliations
  • University of York (GB)
  • European Molecular Biology Laboratory (DE)
  • University of Cambridge (GB)
  • University of Pittsburgh (US)
Corresponding author
  • Hawkins, Dorothy E.D.P.
  • Antson, Alfred A.
ORCID
Venue
Nucleic Acids Research
Publisher
openRxiv
Published
2023-02-24
Volume
51
Issue
13
Pages
7025-7035
ISSN
0305-1048 (ISSN-L)
DOI
10.1101/2023.02.24.529869
PMC
PMC10359639
OpenAlex
W4379966877
Cited by
32
References
1
Citations by year
2026: 11 · 2025: 8 · 2024: 11 · 2023: 2
FWCI
5.17
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Nucleic Acids Research (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (submittedVersion, other-oa)
  • Apollo (University of Cambridge) (submittedVersion, cc-by)
Funders
  • Wellcome Trust (206161, 206377, 095024MA, 224665, 206377/Z/17/Z)
  • Diamond Light Source
  • National Institutes of Health
  • University of California, San Francisco
  • Medical Research Council
  • Biotechnology and Biological Sciences Research Council (AC014501)
  • National Institute of General Medical Sciences (R01GM129325)
  • National Institute of Allergy and Infectious Diseases
Topics
  • Life Sciences › Immunology and Microbiology › Virology › Bacteriophages and microbial interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Enzyme Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
Keywords
bacteriophage HK97, terminase, DNA packaging motor, cos site, portal protein, procapsid, DNA translocation, headful packaging, double-stranded DNA viruses, cryo-EM structure
MeSH terms
Bacteriophages, Capsid, DNA, Viral, Endodeoxyribonucleases, Virus Assembly, Capsid Proteins, DNA Packaging

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