A Cut above the Rest: Characterization of the Assembly of a Large Viral Icosahedral Capsid

Erin R. Reilly, Milky K. Abajorga, Cory Kiser, Nurul Humaira Mohd Redzuan, Zein Haidar, Lily E. Adams, Randy Diaz, Juliana A. Pinzon, André O. Hudson, Lindsay W. Black, Ru Ching Hsia, Susan T. Weintraub, Julie A. Thomas

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The head of Salmonella virus SPN3US is composed of ∼50 different proteins and is unusual because within its packaged genome there is a mass (>40 MDa) of ejection or E proteins that enter the Salmonella cell. The assembly mechanisms of this complex structure are poorly understood. Previous studies showed that eight proteins in the mature SPN3US head had been cleaved by the prohead protease. In this study, we present the characterization of SPN3US prohead protease mutants using transmission electron microscopy and mass spectrometry. In the absence of the prohead protease, SPN3US head formation was severely impeded and proheads accumulated on the Salmonella inner membrane. This impediment is indicative of proteolysis being necessary for the release and subsequent DNA packaging of proheads in the wild-type phage. Proteomic analyses of gp245- proheads that the normal proteolytic processing of head proteins had not occurred. Assays of a recombinant, truncated form of the protease found it was active, leading us to hypothesize that the C-terminal propeptide has a role in targeting the protease into the prohead core. Our findings provide new evidence regarding the essential role of proteolysis for correct head assembly in this remarkable parasite.

Original languageEnglish (US)
Article number725
JournalViruses
Volume12
Issue number7
DOIs
StatePublished - Jul 2020

Keywords

  • Salmonella
  • giant phage
  • head assembly
  • mass spectrometry
  • myovirus
  • prohead protease
  • proteolytic maturation
  • transmission electron microscopy

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

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