Dissecting the herpesvirus architecture by targeted proteolysis

Gina R. Daniel, Caitlin E. Pegg, Gregory A. Smith*

*Corresponding author for this work

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Herpesvirus particles have a complex architecture consisting of an icosahedral capsid that is surrounded by a lipid envelope. Connecting these two components is a layer of tegument that consists of various amounts of 20 or more proteins. The arrangement of proteins within the tegument cannot easily be assessed and instead is inferred from tegument interactions identified in reductionist models. To better understand the tegument architecture, we have developed an approach to probe capsid-tegument interactions of extracellular viral particles by encoding tobacco etch virus (TEV) protease sites in viral structural proteins, along with distinct fluorescent tags in capsid and tegument components. In this study, TEV sites were engineered within the pUL36 large tegument protein, a critical structural element that is anchored directly on the capsid surface. Purified pseudorabies virus extracellular particles were permeabilized, and TEV protease was added to selectively cleave the exposed pUL36 backbone. Interactions with the capsid were assessed in situ by monitoring the fate of the fluorescent signals following cleavage. Although several regions of pUL36 are proposed to bind capsids, pUL36 was found stably anchored to the capsid exclusively at its carboxyl terminus. Two additional tegument proteins, pUL37 and pUS3, were tethered to the capsid via pUL36, whereas the pUL16, pUL47, pUL48, and pUL49 tegument proteins were not stably bound to the capsid.

Original languageEnglish (US)
Article numbere00738-18
JournalJournal of virology
Volume92
Issue number17
DOIs
StatePublished - Sep 1 2018

Keywords

  • Capsid
  • Herpesvirus
  • PRV
  • Pseudorabies virus
  • Selective disassembly
  • Structure
  • TEV protease
  • Tegument
  • Virion

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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