A mutation in the ectodomain of herpes simplex virus 1 glycoprotein B causes defective processing and retention in the endoplasmic reticulum.
Herpes simplex virus 1 (HSV-1) glycoprotein B (gB) is one of several envelope glycoproteins required for virion infectivity and is the only one known to oligomerize into homodimers. To study the conformational constraints for translocation of HSV-1 gB to the surface of eukaryotic cells, we analyzed the transport through the exocytic pathway of the wild-type glycoprotein and of mutant forms with insertions in the ectodomain and intracellular carboxy terminus. Transient expression of the glycoproteins in COS-1 cells showed that an insertion at position 479 in the amino-terminal ectodomain of gB, shown previously by reactions with monoclonal antibodies to have altered the conformation of the molecule, also had a drastic effect on transport, precluding exit of the mutant from the endoplasmic reticulum (ER) and transport to the Golgi and the plasma membrane. The fact that the mutant, gB-(Lk479), formed dimers suggests that local changes in assembled regions caused the transport defect. Mutants containing insertions at residues 600 of the ectodomain and 810 in the intracellular domain were slightly retarded in their rate of transport from the ER to the Golgi. The glucose-regulated proteins GRP78 and GRP94, which are resident proteins of the ER, associated with partially glycosylated, faster-migrating forms of gB but not with the fully processed, more slowly migrating product. GRP78 and GRP94 formed complexes with the mutant gB-(Lk479), which was degraded in the ER. Our results indicate that GRP78, and perhaps also GRP94, acts as a chaperone in the assembly of native gB oligomers and also binds to aberrant forms of the molecule, arresting their transport from the ER and possibly serving as markers for protein degradation in this compartment of the exocytic pathway.
Navarro
D
D
Division of Oral Biology, School of Dentistry, University of California, San Francisco 94143-0512.
Qadri
I
I
Pereira
L
L
D000818:Animals; D000911:Antibodies, Monoclonal; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000502:physiology*; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D018139:Simplexvirus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014771:Virion / Q000235:genetics / Q000502:physiology
D000911:Antibodies, Monoclonal; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone