The effects of packaged, but misguided, single-stranded DNA genomes are transmitted to the outer surface of the f X174 capsid

Elizabeth T. Ogunbunmi, Aaron P. Roznowski, Bentley A. Fane

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Most icosahedral viruses condense their genomes into volumetrically constrained capsids. However, concurrent genome biosynthesis and packaging are specific to single-stranded DNA (ssDNA) viruses. ssDNA genome packaging combines elements found in both double-stranded DNA (dsDNA) and ssRNA systems. Similar to dsDNA viruses, the genome is packaged into a preformed capsid. Like ssRNA viruses, there are numerous capsid-genome associations. In ssDNA microviruses, the DNA-binding protein J guides the genome between 60 icosahedrally ordered DNA binding pockets. It also partially neutralizes the DNA’s negative phosphate backbone. f X174-related microviruses, such as G4 and a3, have J proteins that differ in length and charge organization. This suggests that interchanging J proteins could alter the path used to guide DNA in the capsid. Previously, a f XG4J chimera, in which the f X174 J gene was replaced with the G4 gene, was characterized. It displayed lethal packaging defects, which resulted in procapsids being removed from productive assembly. Here, we report the characterization of another inviable chimera, f Xa3J. Unlike f XG4J, f Xa3J efficiently packaged DNA but produced noninfectious particles. These particles displayed a reduced ability to attach to host cells, suggesting that internal DNA organization could distort the capsid’s outer surface. Mutations that restored viability altered J-coat protein contact sites. These results provide evidence that the organization of ssDNA can affect both packaging and postpackaging phenomena. IMPORTANCE ssDNA viruses utilize icosahedrally ordered protein-nucleic acids interactions to guide and organize their genomes into preformed shells. As previously demonstrated, chaotic genome-capsid associations can inhibit f X174 packaging by destabilizing packaging complexes. However, the consequences of poorly organized genomes may extend beyond the packaging reaction. As demonstrated herein, it can lead to uninfectious packaged particles. Thus, ssDNA genomes should be considered an integral and structural virion component, affecting the properties of the entire particle, which includes the capsid’s outer surface.

Original languageEnglish (US)
Article numbere00883-21
JournalJournal of virology
Issue number18
StatePublished - Sep 2021


  • Bacteriophage
  • DNA packaging
  • F X174
  • Single-stranded DNA
  • Virus assembly

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology


Dive into the research topics of 'The effects of packaged, but misguided, single-stranded DNA genomes are transmitted to the outer surface of the f X174 capsid'. Together they form a unique fingerprint.

Cite this