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Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
08:31

Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'

Published on: May 26, 2013

Periodic genome sequences facilitate packaging in a single-stranded DNA virus.

Elizabeth T Ogunbunmi1,2, Megan Mokriski3, April D Burch3

  • 1The BIO5 Institute, University of Arizona, Tucson, Arizona, USA.

Journal of Virology
|July 1, 2026
PubMed
Summary
This summary is machine-generated.

Single-stranded DNA (ssDNA) viruses package their genomes into capsids. Altering specific sequence motifs in the øX174 virus genome disrupts this packaging process, revealing insights into viral genome organization.

Keywords:
DNA packagingMicroviridaephiX174

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Area of Science:

  • Virology
  • Molecular Biology
  • Structural Biology

Background:

  • Icosahedral viruses package genomes into constrained capsids.
  • Single-stranded DNA (ssDNA) viruses like øX174 package genomes concurrently with replication into preformed capsids.
  • The mechanism of ssDNA genome accommodation within capsids is not fully understood.

Purpose of the Study:

  • To investigate the role of sequence motifs in the øX174 genome during packaging.
  • To elucidate the ssDNA viral DNA replication-packaging pathway.

Main Methods:

  • Site-directed mutagenesis was used to alter sequence motifs in the øX174 genome, creating the mutant øXDO5.
  • Cells were co-infected with wild-type and øXDO5 viruses to track genome replication and packaging.
  • Comparative analysis of viral DNA replication and virion content was performed.

Main Results:

  • Mutating sequence motifs in the øXDO5 genome resulted in packaging defects.
  • Replication of øXDO5 genomes was comparable to wild-type early in the infection.
  • Significantly fewer øXDO5 genomes were found within mature virions compared to wild-type.

Conclusions:

  • Periodic sequence motifs in the øX174 genome are crucial for efficient viral genome packaging.
  • These findings suggest selective pressure on ssDNA viral genomes to maintain specific packaging-related sequence patterns.
  • The study highlights a defect in the transition from packaging intermediates to mature virions for the øXDO5 mutant.