Characterization of a 52K protein of murine cytomegalovirus and its immunological cross-reactivity with the DNA-binding protein ICP36 of human cytomegalovirus
We have developed a hybridoma, designated 25G11, which produced a monoclonal antibody (MAb) reactive with a 52K protein of murine cytomegalovirus (MCMV). This MAb, 25G11, was reactive with a protein band of 52K in MCMV-infected cell lysates and with a protein of 49K in human CMV (HCMV)-infected cell lysates as detected by immunoblot analysis. With purified MCMV virions, 25G11 gave a faintly immunoreactive band of 52K. However, no immunoreactive protein band was detected with purified HCMV virions, nor with purified HCMV or MCMV envelope preparations. By immunocytochemistry, 25G11 detected viral antigen primarily in the nucleus of HCMV- or MCMV-infected cells. The antibody 25G11 was used to screen a λgt11 library of HCMV DNA fragments. One of the isolated clones (λ32323B) was employed for gene mapping on the HCMV genome, which suggested that the immunoreactive HCMV protein was the DNA-binding protein (ICP36). Analysis of the recombinant fusion protein with antibody 25G11 and with an MAb (CH16) specific for an HCMV DNA-binding protein confirmed the identity of the cross-reacting protein as ICP36. Furthermore, we found that whereas the epitope recognized by 25G11 was conserved between HCMV and MCMV proteins, the epitope recognized by CH16 was unique to HCMV and thus represents a variable region in the protein.
AndersD. G.1990; Nucleotide sequence of a cytomegalovirus single-stranded DNA-binding protein gene: comparison with alpha- and gammaherpesvirus counterparts reveals conserved segments. Journal of General Virology 71:2451–2456
AndersD. G.,
KiddJ. R.,
GibsonW.1987; Immunological characterization of an early cytomegalovirus single-stranded DNA-binding protein with similarities to the HSV major DNA-binding protein. Virology 161:579–588
CheeM. S.,
BankierA. T.,
BeckS.,
BohniR.,
BrownC. M.,
CernyR.,
HorsnellT.,
HutchisonC. A.III,
KouzaridesT.,
MartignettiE.,
PreddieE.,
SatchwellS. C.,
TomlinsonP.,
WestonK. M.,
BarrellB. G.1990; Analysis of the proteincoding content of the sequence of human cytomegalovirus strain AD169. Current Topics in Microbiology and Immunology 154:125–169
CranageP. M.,
KouzaridesT.,
BankierA. T.,
SatchwellS.,
WestonK.,
TomlinsonP.,
BarrellB. G.,
HartH.,
BellS. E.,
MinsonA. C,
SmithG. L.1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO Journal 5:3057–3063
CranageM. P.,
SmithG. L.,
BellS. E.,
HartH.,
BrownC.,
BankierA. T.,
TomlinsonP.,
BarrellB. G.,
MinsonA. C.1988; Identification and expression of a human cytomegalovirus glycoprotein with homology to Epstein-Barr virus BXLF2 product, varicella-zoster virus gpIII, and herpes simplex virus type 1 glycoprotein H. Journal of Virology 62:1416–1422
KembleG. W.,
McCormickA. L.,
PereiraL.,
MocarskiE. S.1987; A cytomegalovirus protein with properties of herpes simplex virus ICP8: partial purification of the polypeptide and map position of the gene. Journal of Virology 61:3143–3151
LeachF. S.,
MocarskiE. S.1989; Regulation of cytomegalovirus late-gene expression: differential use of three start sites in the transcriptional activation of ICP36 gene expression. Journal of Virology 63:1783–1791
LohL. C.,
BalachandranN.,
QualtiereL. F.1988; Characterization of a major virion envelope glycoprotein complex of murine cytomegalovirus and its immunological cross-reactivity with human cytomegalovirus. Virology 166:206–216
MeijerH.,
DormansP. H. J.,
GeelenJ. L. M. C.,
Van BovenC. P. A.1984; Rat cytomegalovirus: studies on the viral genome and the proteins of virions and nucleocapsids. Journal of General Virology 65:681–695
MocarskiE. S.,
PereiraL.,
MichaelN.1985; Precise localization of genes on large animal virus genomes: use of λ gtl1 and monoclonal antibodies to map the gene for a cytomegalovirus protein family. Proceedings of the National Academy of Sciences, U.S.A 82:1266–1270
PandeH.,
CampoK.,
ChurchillM. A.,
ClarkB. R.,
ZaiaJ. A.1988; Genomic localization of a 32-kDa capsid protein of human cytomegalovirus. Virology 167:306–310
PandeH.,
CampoK.,
ChurchillM. A.,
ZaiaJ. A.1989; Genomic locus for a 140 kDa structural protein (ppl50) of human cytomegalovirus in strains Towne and AD169. Virus Research 12:11–18
PereiraL.,
HoffmanM.,
GalloD.,
CremerN.1982; Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants. Infection and Immunity 36:924–932
Characterization of a 52K protein of murine cytomegalovirus and its immunological cross-reactivity with the DNA-binding protein ICP36 of human cytomegalovirus