Function of herpes simplex virus type 1 gD mutants with different receptor-binding affinities in virus entry and fusion.
We have studied the receptor-specific function of four linker-insertion mutants of herpes simplex virus type 1 glycoprotein D (gD) representing each of the functional regions of gD. We used biosensor analysis to measure binding of the gD mutants to the receptors HVEM (HveA) and nectin-1 (HveC). One of the mutants, gD(inverted Delta 34t), failed to bind HVEMt but showed essentially wild-type (WT) affinity for nectin-1t. The receptor-binding kinetics and affinities of the other three gD mutants varied over a 1,000-fold range, but each mutant had the same affinity for both receptors. All of the mutants were functionally impaired in virus entry and cell fusion, and the levels of activity were strikingly similar in these two assays. gD(inverted Delta 34)-containing virus was defective on HVEM-expressing cells but did enter nectin-1-expressing cells to about 60% of WT levels. This showed that the defect of this form of gD on HVEM-expressing cells was primarily one of binding and that this was separable from its later function in virus entry. gD(inverted Delta 243t) showed WT binding affinity for both receptors, but virus containing this form of gD had a markedly reduced rate of entry, suggesting that gD(inverted Delta 243) is impaired in a postbinding step in the entry process. There was no correlation between gD mutant activity in fusion or virus entry and receptor-binding affinity. We conclude that gD functions in virus entry and cell fusion regardless of its receptor-binding kinetics and that as long as binding to a functional receptor occurs, entry will progress.
Milne
Richard S B
RS
Department of Microbiology and Center for Oral Health Research, School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6002, USA. rmilne@biochem.dental.upenn.edu
Hanna
Sheri L
SL
Rux
Ann H
AH
Willis
Sharon H
SH
Cohen
Gary H
GH
Eisenberg
Roselyn J
RJ
D002460:Cell Line; D005816:Genetic Complementation Test; D018259:Herpesvirus 1, Human / Q000502:physiology*; D008561:Membrane Fusion / Q000502:physiology*; D008958:Models, Molecular; D011991:Receptors, Virus / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D011991:Receptors, Virus; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1