Bovine herpesvirus 1 UL49.5 homolog gene encodes a novel viral envelope protein that forms a disulfide-linked complex with a second virion structural protein.
We previously reported that the genome of bovine herpesvirus 1 (BHV-1) contains an open reading frame (ORF) homologous to the herpes simplex virus UL49.5 ORF, and as with the herpes simplex virus UL49.5 ORF, the deduced amino acid sequence of the BHV-1 UL49.5 homolog (UL49.5h) contains features characteristic of an integral membrane protein, implying that it may constitute a functional gene encoding a novel viral envelope protein. This communication reports on the identification of the BHV-1 UL49.5h gene product. By employing an antibody against a synthetic BHV-1 UL49.5h peptide and an UL49.5h gene deletion mutant, the primary product of BHV-UL49.5h gene was identified as a polypeptide with a size of approximately 9 kDa; in both infected cells and isolated virions, the UL49.5h products were found to exist in three forms; monomer, disulfide-linked homodimer, and disulfide-linked heterodimer containing a second viral protein with a size of about 39 kDa. O-Glycosidase digestion and [3H]glucosamine labelling experiments showed that the UL49.5h protein is not glycosylated. Although the deduced amino acid sequence contains putative sites for myristylation and phosphorylation, we were unable to detect either modification. Surface labelling and trypsin digestion protection experiments showed that the BHV-1 UL49.5h protein was present on the surface of infected cells and on the surface of mature virions. Nonionic detergent partition of isolated virions revealed that the UL49.5h protein is more tightly associated with the virion tegument-nucleocapsid structure than envelope protein gD. The results from this study demonstrate that the BHV-1 UL49.5h gene encodes a nonglycosylated virion envelope protein which may associate with virion internal structures by forming a complex with the 39-kDa virion structural protein.
Liang
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X
Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Canada.
Chow
B
B
Raggo
C
C
Babiuk
L A
LA
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D005814:Genes, Viral; D006031:Glycosylation; D007242:Herpesvirus 1, Bovine / Q000737:chemistry / Q000235:genetics*; D009227:Myristic Acids; D017830:Octoxynol; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D010766:Phosphorylation; D011092:Polyethylene Glycols / Q000494:pharmacology; D011817:Rabbits; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000737:chemistry; D014771:Virion / Q000187:drug effects
D004220:Disulfides; D009227:Myristic Acids; D010455:Peptides; C099455:UL49.5 protein, bovine herpesvirus type 1; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40