The UL34 gene of herpes simplex virus type 2 (HSV-2) is highly conserved in the herpesvirus family. The UL34 gene product was identified In lysates of HSV-2-infected cells as protein species with molecular masses of 31 and 32·5 kDa, the latter being a phosphorylated product. Synthesis of these proteins occurred at late times post-infection and was highly dependent on viral DNA synthesis. Immunofluorescence assays revealed that the UL34 protein was localized in the cytoplasm in a continuous net-like structure, closely resembling the staining pattern of the endoplasmic reticulum (ER), in both HSV-2-infected cells and in cells transiently expressing UL34 protein. Deletion mutant analysis showed that this colocalization required the C terminus of the UL34 protein. The UL34 protein associated with virions but not with A, B or C capsids. We treated virions, HSV-2-infected cells and cells expressing the UL34 protein with a protease in order to examine the topology of the UL34 protein. In addition, we constructed UL34 deletion mutant proteins and examined their intracellular localization. Our data strongly support the hypothesis that the UL34 protein is inserted into the viral envelope as a tail-anchored type II membrane protein and is significant for virus envelopment.
AlbrechtJ. C.,
NicholasJ.,
BillerD.,
CameronK. R.,
BiesingerB.,
NewmanC.,
WittmannS.,
CraxtonM. A.,
ColemanH.,
FleckensteinB.1992; Primary structure of the herpesvirus saimiri genome. Journal of Virology 66:5047–5058
BrideauA. D.,
BanfieldB. W.,
EnquistL. W.1998; The Us9 gene product of pseudorabies virus, an alphaherpesvirus, is a phosphorylated, tail-anchored type II membrane protein. Journal of Virology 72:4560–4570
CheeM. S.,
BankierA. T.,
BeckS.,
BohniR.,
BrownC. M.,
ChernyR.,
HorsnellT.,
HutchinsonC. A.III.,
KouzaridesT.,
MartignettiJ. A.,
PreddieE.,
SatchwellS. C.,
TomlinsonP.,
WestonK. M.,
BarrellB. G.1990; Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD 169. Current Topics in Microbiology and Immunology 154:125–169
DaikokuT.,
YamashitaY.,
TsurumiT.,
MaenoK.,
NishiyamaY.1993; Purification and biochemical characterization of the protein kinase encoded by the US3 gene of herpes simplex virus type 2. Journal of General Virology 73:303–311
DaikokuT.,
KurachiR.,
TsurumiT.,
NishiyamaY.1994; Identification of a target protein of US3 kinase of herpes simplex virus type 2. Journal of General Virology 75:2065–2068
DolanA.,
JamiesonF. E.,
CunninghamC.,
BarnettB. C.,
McGeochD. J.1998; The genome sequence of herpes simplex virus type 2. Journal of Virology 72:2010–2021
FrangioniJ. V.,
BeahmP. H.,
ShifrinV.,
JostC. A.,
NeelB. G.1992; The nontransmembrane tyrosine phosphatase PTP-1B localizes to the endoplasmic reticulum via its 35 amino acid C-terminal sequence. Cell 68:545–560
GompelsU. A.,
NicholasJ.,
LawrenceG.,
JonesM.,
ThomsonB. J.,
MartinM. E. D.,
EfstathiouS.,
CraxtonM.,
MacaulayH. A.1995; The DNA sequence of human herpesvirus-6: structure, coding content, and genome evolution. Virology 209:29–51
LinstedtA. D.,
FoguetM.,
RenzM.,
SeeligH. P.,
GlickB. S.,
HauriH. P.1995; A C-terminally anchored Golgi protein is inserted into the endoplasmic reticulum and then transported to the Golgi apparatus. Proceedings of the National Academy of Sciences, USA 92:5102–5105
LukasJ. R.,
AignerM.,
DenkM.,
HeinzlH.,
BuriunM.,
MayrR.1998; Carbocyanine postmortem neuronal tracing: influence of different parameters on tracing distance c with immunocytochemistry. Journal of Histochemistry and Cytochemistry 46:901–910
NishiyamaY.,
YamadaY.,
KurachiR.,
DaikokuT.1992; Construction of a US3 lacZ insertion mutant of herpes simplex virus type 2 and characterization of its phenotype in vitro and in vivo. Virology 190:256–268
PfitznerA. J.,
TsaiE. C.,
StromingerJ. L.,
SpeckS. H.1987; Isolation and characterization of cDNA clones corresponding to transcripts from the Bam HI H and F region of the Epstein–Barr virus genome. Journal of Virology 61:2902–2909
PurvesF. C.,
SpectorD.,
RoizmanB.1991; The herpes simplex virus 1 protein kinase encoded by the US3 gene mediates posttranslational modification of the phosphoprotein encoded by the UL34 gene. Journal of Virology 65:5757–5764
PurvesF. C.,
SpectorD.,
RoizmanB.1992; UL34, the target of the herpes simplex virus US3 protein kinase, is a membrane protein which in its unphosphorylated state associates with novel phosphoproteins. Journal of Virology 66:4295–4303
RoizmanB.1996; The function of herpes simplex virus genes: a primer for genetic engineering of novel vectors. Proceedings of the National Academy of Sciences USA: 93:11307–11312
RoizmanB.,
SearsA. E.1996; Herpes simplex viruses and their replication. InFields Virology pp 1043–1107 Edited by
FieldsB. N.,
KnipeD. M.,
HowleyP. M.
Philadelphia: Lippincott–Raven;
RollerR. J.,
RoizmanB.1991; Herpes simplex virus 1 RNA binding protein US11 negatively regulates the accumulation of a truncated viral mRNA. Journal of Virology 65:5873–5879
StackpoleC. W.1969; Herpes-type virus of the frog renal adenocarcinoma. I. Virus development in tumor transplants maintained at low temperature. Journal of Virology 4:75–93
TerasakiM.,
JaffeL. A.1991; Organization of the sea urchin egg endoplasmic reticulum and its reorganization at fertilization. Journal of Cell Biology 114:929–940
TerasakiM.,
SongJ. D.,
WongJ. R.,
WeissM. J.,
ChenL. B.1984; Localization of endoplasmic reticulum in living and glutaraldehyde fixed cells with fluorescent dyes. Cell 38:101–108
WhealyM. E.,
CarsJ. P.,
MeadeR. P.,
RobbinsA. K.,
EnquistL. W.1991; Effect of brefeldin A on alphaherpesvirus membrane protein glycosylation and virus egress. Journal of Virology 65:1066–1081
WhitleyP.,
GrahnE.,
KutayU.,
RapoportT. A.,
von HeijneG.1996; A 12-residue-long polyleucine tail is sufficient to anchor synaptobrevin to the endoplasmic reticulum membrane. Journal of Biological Chemistry 271:7583–7586
YeG. J.,
VaughanK. T.,
ValleeR. B.,
RoizmanB.2000; The herpes simplex virus 1 UL34 protein interacts with a cytoplasmic dynein intermediate chain and targets nuclear membrane. Journal of Virology 74:1355–1363