Herpes simplex virus glycoprotein B mutations define structural sites in domain I, the membrane proximal region, and the cytodomain that regulate entry

Qing Fan, Richard Longnecker, Sarah A. Connolly*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

The viral fusion protein glycoprotein B (gB) is conserved in all herpesviruses and is essential for virus entry. During entry, gB fuses viral and host cell membranes by refolding from a prefusion to a postfusion form. We previously introduced three structure-based mutations (gB-I671A/H681A/F683A) into the domain V arm of the gB ectodomain that resulted in reduced cell-cell fusion. A virus carrying these three mutations (called gB3A) displayed a small-plaque phenotype and remarkably delayed entry into cells. To identify mutations that could counteract this phenotype, we serially passaged the gB3A virus and selected for revertant viruses with increased plaque sizes. Genomic sequencing revealed that the revertant viruses had second-site mutations in gB, including E187A, M742T, and S383F/G645R/V705I/V880G. Using expression constructs encoding these mutations, only gB-V880G was shown to enhance cell-cell fusion. In contrast, all of the revertant viruses showed enhanced entry kinetics, underscoring the fact that cell-cell fusion and virus-cell fusion are different. The results indicate that mutations in three different regions of gB (domain I, the membrane proximal region, and the cytoplasmic tail domain) can counteract the slow-entry phenotype of gB3A virus. Mapping these compensatory mutations to prefusion and postfusion structural models suggests sites of intramolecular functional interactions with the gB domain V arm that may contribute to the gB fusion function.

Original languageEnglish (US)
Article numbere01050-21
JournalJournal of virology
Volume95
Issue number22
DOIs
StatePublished - Nov 2021

Keywords

  • Entry
  • Fusion
  • Glycoprotein gB
  • Glycoproteins
  • Herpes simplex virus 1
  • Revertant

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Fingerprint

Dive into the research topics of 'Herpes simplex virus glycoprotein B mutations define structural sites in domain I, the membrane proximal region, and the cytodomain that regulate entry'. Together they form a unique fingerprint.

Cite this