Abstract
The tegument phosphoproteins of human cytomegalovirus (HCMV) elicit cytotoxic T-lymphocyte (CTL) responses and are hence candidates for subunit vaccine development. Little is known, however, about the tegument proteins of nonhuman cytomegaloviruses, such as guinea pig CMV (GPCMV). DNA sequence analysis of the Eco R I “C” fragment of the GPCMV genome identified an open reading frame (ORF) which is colinear with that of the HCMV tegument phosphoprotein, UL83 (pp65). This ORF was found to have identity to HCMV UL83 and was predicted to encode a 565-amino-acid (aa) protein with a molecular mass of 62.3 kDa. Transcriptional analyses revealed that a GPCMV UL83 probe hybridized with both 2.2 kb and 4.2 kb mRNA species at 48 h post-infection (p.i.); synthesis of these messages was blocked by phosphonoacetic acid (PAA), defining these as “late” gene transcripts. In vitro translation of the UL83 ORF in reticulocyte lysate resulted in synthesis of a 65 kDa protein. Immunofluorescence experiments revealed that the putative GPCMV UL83 homolog exhibited a predominantly nuclear localization pattern. Polyclonal antisera were raised against a UL83/glutathione-S-transferase (GST) fusion protein. This antibody identified a 70-kDa virion-associated protein, the putative GPCMV UL83 homolog, in immunoblot and radioimmunoprecipitation experiments. Labeling experiments with 32P-orthophosphate indicated that the GPCMV UL83 protein is phosphorylated. Western blot analysis of glycerol tartrate gradient-purified virions and dense bodies confirmed that the putative GPCMV UL83 homolog was a constituent of both fractions.
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Alford C.A. and BrittW.J., in: Fields B.N. and Knipe D.M. (ed), Virology second edition. Raven Press, New York, 1981±2010, 1990.
Boppana S.B., Pass R.F., Britt W.J., Stagno S., and Alford C.A., Pediatr Infect Dis J 11, 93±99, 1992.
Fowler K.B., Stagno S., Pass R.F., Britt W.J., Boll T.J., and Alford C.A., N Engl J Med 326, 663±667, 1992.
Grif®th B.P. and Aquino-De Jesus M.J.C., Transplant Proc 23, 29±31, 1991.
Grif®th B.P., McCormick S.R., Fong C.K.Y., Lavallee J.T., Lucia H.L., and Goff E., J Virol 55, 402±409, 1985.
Bratcher D.F., Bourne N., Bravo F.J., Schleiss M.R., Slaoui M., Myers M.G., and Bernstein D.I., J Infect Dis 172, 944±950, 1995.
Schleiss M.R., Bourne N., and Bernstein D.I., Pediatr Res 43, 252A, 1998.
Schleiss M.R., Virology 202, 173±185, 1994.
Brady R.C. and Schleiss M.R., Arch Virol 141, 2409±2424, 1996.
Spaete R.R., Gehrz R.C., and Landini M.P., J Gen Virol 75, 3287±3308, 1994.
McLaughlin-Taylor E., Pande H., Forman S.J., Tanamachi B., Li C.R., Zaia J.A., Greenberg P.D., and Riddell S.R., J Med Virol 43, 103±110, 1994.
Wills M.R., Carmichael A.J., Mynard K., Jin X., Weekes M.P., Plachter B., and Sissons J.G., J Virol 70, 7569±7579, 1996.
Gilbert M.J., Riddell S.R., Plachter B., and Greenberg P.D., Nature 383, 720±722, 1996.
Walter E.A., Greenberg P.D., Gilbert M.J., Finch R.J., Watanabe K.S., Thomas E.D., and Riddell S.R., N Engl J Med 333, 1038±1044, 1995.
Bishopp F., Bioworld Today 7(231), 1, 4, 1996.
Gao M. and Isom H.C., J Virol 52, 436±447, 1984.
Maniatis T., Fritsch E.F., and Sambrook J., Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1982.
Yin C.-Y., Gao M., and Isom H.C., J Virol 64, 1537±1548, 1990.
Rixon F.J., Addison C., McGregor A., Macnab S.J., Nicholson P., Preston V.G., and Tatman J.D., J Gen Virol 77, 2251±2260, 1996.
Smith D.B. and Johnson K.S., Gene 67, 31±40, 1988.
Paglino J.C., Brady R.C., and Schleiss M.R., Arch Virol 144, 447±462, 1999.
Laemmli U.K., Nature 227, 680±685, 1970.
Talbot P. and Almeida J.D., J Gen Virol 36, 345±349, 1977.
Schleiss M.R., Aktan L., and Erdem G., Abstracts of the XXII International Hervesvirus Workshop, Abstract #434, 1997.
Isom H.C., Gao M., and Wigdahl B., J Virol 49, 426±436, 1984.
McLauchlan J., Gaffney D., Whitton J.L., and Clements J.B., Nucleic Acids Res 13, 1347±1366, 1985.
Chee M.S., Bankier A.T., Beck S., Bohni R., Brown C.M., Cerney R., Horsnell T., Hutchinson C.A., Kouzarides T., Martignetti J.A., Preddie E., Satchwell S.C., Tomlinson P., Weston K.M., and Barrell B.G., Curr Top Microbiol Immunol 154, 125±169, 1990.
RuÈger B., Klages S., Walla B., Albrecht J., Fleckenstein B., Tomlinson P., and Barrell B., J Virol 61, 446±453, 1987.
Cranmer L.D., Clark C.L., Morello C.S., Farrell H.E., Rawlinson W.D., and Spector D.H., J Virol 70, 7929±7939, 1996.
Schmolke S., Drescher P., Jahn G., and Plachter B., J Virol 69, 1071±1078, 1995.
Gallina A., Percivalle E., Simoncini L., Revello M.G., Gerna G., and Milanesi G., J Gen Virol 77, 1151±1157, 1996.
Faisst S. and Meyer S., Nucleic Acids Res 20, 3±26, 1992.
Depto A.S. and Stenberg R.M., J Virol 63, 1232±1238, 1989.
Geballe A.P., Leach F.S., and Mocarski E.S., J Virol 57, 864± 874, 1986.
Pande H., Lee T.D., Churchill M.A., and Zaia J.A., Virology 178, 6±14.
Michelson S., Tardy-Panit M., and Barzu O., J Biochem 149, 393±399, 1985.
Somogyi T., Michelson S., and Masse M.J., Virology 174, 276± 285, 1990.
Gallina A., Simoncini L., Garbelli S., Percivalle E., Pedrali-Noy G., Lee K.S., Erikson R.L., Plachter B., Gerna G., and Milanesi G., J Virol 73, 1468±1478, 1999.
Schleiss M., Bourne N., Jensen N., Bravo F., Reece A., Erdem G., and Bernstein D., Pediatr Res 45, 173A, 1999.
Pande H., Campo K., Tanamachi B., Forman S.J., and Zaia J.A., Scand J Infect Dis (Suppl. 99), 117±120, 1995.
Sanchez V., Angeletti P.C., Engler J.A., and Britt W.J., J Virol 72, 3321±3329.
Dal Monte P., Bessia C., Landini M.P., and Michelson S., J Gen Virol 77, 2591±2596, 1996.
McGregor A. and Schleiss M.R., unpublished data.
Schmolke S., Kern H.F., Drescher P., Jahn G., and Plachter B., J Virol 69, 5959±5968, 1995.
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Schleiss, M.R., McGregor, A., Jensen, N.J. et al. Molecular Characterization of the Guinea Pig Cytomegalovirus UL83 (pp65) Protein Homolog. Virus Genes 19, 205–221 (1999). https://doi.org/10.1023/A:1008136714136
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DOI: https://doi.org/10.1023/A:1008136714136