Herpes simplex virus type 1 mediates fusion through a hemifusion intermediate by sequential activity of glycoproteins D, H, L, and B

Proc Natl Acad Sci U S A. 2007 Feb 20;104(8):2903-8. doi: 10.1073/pnas.0608374104. Epub 2007 Feb 13.

Abstract

Virus-induced membrane fusion can be subdivided into three phases defined by studies of class I and class II fusion proteins. During Phase I, two membranes are brought into close apposition. Phase II marks the mixing of the outer membrane leaflets leading to formation of a hemifusion intermediate. A fusion pore stably forms and expands in Phase III, thereby completing the fusion process. Herpes simplex virus type 1 (HSV-1) requires four glycoproteins to complete membrane fusion, but none has been defined as class I or II. Therefore, we investigated whether HSV-1-induced membrane fusion occurred following the same general phases as those described for class I and II proteins. In this study we demonstrate that glycoprotein D (gD) and the glycoprotein H and glycoprotein L complex (gHL) mediated lipid mixing indicative of hemifusion. However, content mixing and full fusion required glycoprotein B (gB) to be present along with gD and gHL. Our results indicate that, like class I and II fusion proteins, fusion mediated by HSV-1 glycoproteins occurred through a hemifusion intermediate. In addition, both gB and gHL are probably directly involved in the fusion process. From this, we propose a sequential model for fusion via HSV-1 glycoproteins whereby gD is required for Phase I, gHL is required for Phase II, and gB is required for Phase III. We further propose that glycoprotein H and gB are likely to function sequentially to promote membrane fusion in other herpesviruses such as Epstein-Barr virus and human herpesvirus 8.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Viral / pharmacology
  • CHO Cells
  • Chlorocebus aethiops
  • Cricetinae
  • Cricetulus
  • Glycoproteins / metabolism*
  • Green Fluorescent Proteins / metabolism
  • Herpesvirus 1, Human / drug effects
  • Herpesvirus 1, Human / metabolism*
  • Lipid Metabolism / drug effects
  • Membrane Fusion* / drug effects
  • Mutation / genetics
  • Recombinant Fusion Proteins / metabolism
  • Vero Cells
  • Viral Proteins / metabolism*

Substances

  • Antibodies, Viral
  • Glycoproteins
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Green Fluorescent Proteins