Mutations in the fusion protein of measles virus that confer resistance to the membrane fusion inhibitors carbobenzoxy-D-Phe-L-Phe-Gly and 4-nitro-2- phenylacetyl amino-benzamide

Michael N. Ha, Sébastien Delpeut, Ryan S. Noyce, Gary Sisson, Karen M. Black, Liang Tzung Lin, Darius Bilimoria, Richard K. Plemper, Gilbert G. Privé, Christopher D. Richardson

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The inhibitors carbobenzoxy (Z)-D-Phe-L-Phe-Gly (fusion inhibitor peptide [FIP]) and 4-nitro-2-phenylacetyl amino-benzamide (AS-48) have similar efficacies in blocking membrane fusion and syncytium formation mediated by measles virus (MeV). Other homologues, such as Z-D-Phe, are less effective but may act through the same mechanism. In an attempt to map the site of action of these inhibitors, we generated mutant viruses that were resistant to the inhibitory effects of Z-D-Phe-LPhe- Gly. These 10 mutations were localized to the heptad repeat B (HRB) region of the fusion protein, and no changes were observed in the viral hemagglutinin, which is the receptor attachment protein. Mutations were validated in a luciferase-based membrane fusion assay, using transfected fusion and hemagglutinin expression plasmids or with syncytium-based assays in Vero, Vero-SLAM, and Vero-Nectin 4 cell lines. The changes I452T, D458N, D458G/V459A, N462K, N462H, G464E, and I483R conferred resistance to both FIP and AS-48 without compromising membrane fusion. The inhibitors did not block hemagglutinin protein-mediated binding to the target cell. Edmonston vaccine/laboratory and IC323 wild-type strains were equally affected by the inhibitors. Escape mutations were mapped upon a three-dimensional (3D) structure modeled from the published crystal structure of parainfluenzavirus 5 fusion protein. The most effective mutations were situated in a region located near the base of the globular head and its junction with the alpha-helical stalk of the prefusion protein. We hypothesize that the fusion inhibitors could interfere with the structural changes that occur between the prefusion and postfusion conformations of the fusion protein.

Original languageEnglish
Article numbere01026-17
JournalJournal of Virology
Volume91
Issue number23
DOIs
Publication statusPublished - Dec 1 2017

Keywords

  • Antiviral inhibitors
  • AS-48
  • Drug resistance
  • Escape mutations
  • F protein
  • FIP
  • Fusion inhibitors
  • Measles virus
  • Membrane fusion
  • Z-D-Phe-L-Phe-Gly

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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