Canine herpesvirus ORF2 is a membrane protein modified by N-linked glycosylation.
Canine herpesvirus (CHV) ORF2, located downstream of the glycoprotein C (gC) gene, has homologues with some of the alphaherpesviruses. To characterize CHV OFR2, a recombinant CHV carrying a LacZ gene in the ORF2 locus, and recombinant vaccinia virus expressing ORF2 protein were constructed. Northern blot analysis revealed ORF2 and a gamma2 class late gene, and its protein product was detectable in CHV-infected cells reacted with ORF2 protein antiserum. Tunicamycin and N-glycosidase F treatment revealed that the ORF2 protein was modified by N-linked glycosylation. Fractionation and immune fluorescence analyses of the CHV-infected cells showed the ORF2 as a membrane protein transportable to the surface of infected cells. In vitro, the ORF2 protein did not affect viral replication and cell-to-cell viral spreading. Present findings represent the first evidence pointing to the CHV ORF2 as a membrane protein modified by an N-linked glycosylation.
Department of Global Agricultural Science, Graduate School of Agricultural and Life Science, The University of Tokyo, Japan.
Yoshifumi
Y
Nishikawa
Michiko
M
Kimura
Xuenan
X
Xuan
Levi
L
Makala
Hideyuki
H
Nagasawa
Takeshi
T
Mikami
Haruki
H
Otsuka
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D004285:Dogs; D005260:Female; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D002184:Herpesvirus 1, Canid / Q000235:genetics* / Q000254:growth & development; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D016366:Open Reading Frames; D012367:RNA, Viral; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
D006023:Glycoproteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins