Herpes simplex virus type 1 gene UL4 is predicted to encode a 199-amino-acid protein with a molecular mass of 21·5 kDa. We report here identification of this protein and its localization in the nuclei of infected cells. Antisera raised against oligopeptides corresponding to the C terminus of the predicted UL4 protein were used for identification of a 25 kDa protein as the product of the UL4 gene. This protein was not detected in cells infected with a UL4 defective mutant virus, but was synthesized by coupled in vitro transcription-translation of the UL4 gene. Synthesis of the 25 kDa protein was blocked by phosphonoacetic acid, an inhibitor of DNA synthesis, indicating that the UL4 gene is expressed with γ kinetics. Subcellular fractionation showed the protein to be localized in the nucleus. It was not detected in virions or light particles.
BainesJ. D.,
RoizmanB.1991; The open reading frames UL3, UL4, UL10 and UL16 are dispensable for the replication ofherpes simplex virus 1 in cell culture. Journal of Virology 65:938–944
BirckbichlerP. J.,
PrymeI. F.1973; Fractionation of membrane bound polysomes, free polysomes and nuclei from tissue-cultured cells. European Journal of Biochemistry 33:369–373
BonnerW. M.,
LaskeyR. A.1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
BrownS. M.,
RitchieD. A.,
Subak-SharpeJ. H.1973; Genetic studies with herpes simplex virus type 1. The isolation of temperaturesensitive mutants, their arrangement into complementation groups and recombination analysis leading to a linkage map. Journal of General Virology 18:329–346
CohenG. H.,
DietzcholdB.,
PoncedeLeonM.,
LongD.,
GolubE.,
VarrichioA.,
PereiraL.,
EisenbergR. J.1984; Localization and synthesis of an antigenic determinant of herpes simplex virus glycoprotein D that stimulates the production of neutralizing antibody. Journal of Virology 49:102–108
DignamJ. D.,
RusselM. L.,
RoederR. G.1983; Accurate transcription initiation by RNA polymerase II in soluble extract from isolated mammalian nuclei. Nucleic Acids Research 11:1475–1489
GoodrichL. D.,
RixonF. J.,
ParrisD. S.1989; Kinetics of expression of the gene encoding the 65-kilodalton DNA-binding protein of herpes simplex virus type l. Journal of Virology 62:137–147
JunP. Y.,
StrelowL. I.,
HermanR. C.,
MarsdenH. S.,
EideT.,
HaarrL.,
LeibD. A.1998; The UL4 gene of herpes simplex virus type 1 is dispensable for latency, reactivation and pathogenesis in mice. Journal of General Virology 79:1603–1611
McGeochD. J.,
DolanA.,
DonaldS.,
RixonF. J.1985; Sequence determination and genetic content of the short unique region of the genome of herpes simplex virus type 1. Journal of Molecular Biology 181:1–13
McGeochD. J.,
DolanA.,
DonaldS.,
BrauerD.H.K.
1986; Complete DNA sequence of the short repeat region in the genome of herpes simplex virus type 1. Nucleic Acids Research 14:1727–1745
McGeochD. J.,
DalrympleM. A.,
DavisonA. J.,
DolanA.,
FrameM. C.,
McNabD.,
PerryL. J.,
ScottJ. E.,
TaylorP.1988; The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. Journal of General Virology 69:1531–1574
McGeochD. J.,
CunninghamC.,
McIntyreG.,
DolanA.1991; Comparative sequence analysis of the long unique regions in the genomes of herpes simplex viruses types 1 and 2. Journal of General Virology 72:3057–3075
McLeanG.,
RixonF.,
LangelandN.,
HaarrL.,
MarsdenH.1990; Identification and characterization of the virion protein products of herpes simplex virus type 1 gene UL47. Journal of General Virology 71:2953–2960
MurphyM.,
SchenkP.,
LankinenH. M.,
CrossA. M.,
TaylorP.,
OwsienkaA.,
HopeR. G.,
LudwigH.,
MarsdenH. S.1989; Mapping of epitopes on the 65K DNA binding protein of herpes simplex virus type l. Journal of General Virology 70:2357–2364
SathananthanB.,
RødahlE.,
EkbergT.,
LangelandN.,
HaarrL.1996; Two-dimensional analysis of [35S]methionine labelled and phosphorylated proteins present in virions and light particles of herpes simplex virus type 1, and detection ofpotentially new structural proteins.. Virus Research 46:1–18
SternsdorfT.,
JensenK.,
ZüchnerD.,
WillH.1997; Cellular localization, expression and structure of the nuclear dot protein 52. Journal of Cell Biology 138:435–448
SzilágyiJ. F.,
CunninghamC.1991; Identification and characterization of a novel non-infectious herpes simplex virus-related particle. Journal of General Virology 72:661–668
TamJ. P.1988; Synthetic peptide vaccine design: synthesis and properties of a high density antigenic peptide system. Proceedings of the National Academy of Sciences, USA 85:5409–5413
VlčekČ.,
BenesV.,
LuZ.,
KutishG. F.,
PačesV.,
RockD.,
LetchworthG. J.,
SchwyzerM.1995; Nucleotide sequence analysis of a 30-kb region of the bovine herpesvirus 1 genome which exhibits a colinear gene arrangement with the UL21 to UL4 genes of herpes simplex virus. Virology 210:100–108