1887

Abstract

A rabbit polyclonal antiserum was raised against a recombinant 6×His–UL3 fusion protein expressed in and used to examine the intracellular localization of the UL3 protein of herpes simplex virus type 2 (HSV-2). The antiserum reacted specifically with 31 and 34 kDa proteins in HSV-2 186-infected Vero cells and with 31 and 35 kDa proteins in UL3-expressing COS-7 cells. The UL3 protein localized both in the cytoplasm and in five to ten bright fluorescent granules in the nucleus close to the nuclear membrane at 4 h post-infection (p.i.). These structures became bigger at 5 h p.i. and showed doughnut-like forms at 6 h p.i. In transfected Vero cells, the UL3 protein localized exclusively in the nucleoplasm and specifically in the nucleolus. Five deletion mutants of the UL3 protein were constructed for transfection assays and the results showed that the region containing amino acids 100–164 was important for nucleolar localization. Moreover, green fluorescent protein (GFP)-targetting experiments showed that the region containing amino acids 100–164 was able to transport non-nucleolar GFP to the nucleolus as a fusion protein.

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1999-08-01
2019-10-23
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References

  1. Baines, J. D. & Roizman, B. ( 1991; ). The open reading frames UL3, UL4, UL10, and UL16 are dispensable for the replication of herpes simplex virus 1 in cell culture. Journal of Virology 65, 938-944.
    [Google Scholar]
  2. Borer, R. A., Lehner, C. F., Eppenberger, H. M. & Nigg, E. A. ( 1989; ). Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell 56, 379-390.[CrossRef]
    [Google Scholar]
  3. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W. & Prasher, D. C. ( 1994; ). Green fluorescent protein as a marker for gene expression. Science 263, 802-805.[CrossRef]
    [Google Scholar]
  4. Coleman, N. A. & Peeples, M. E. ( 1993; ). The matrix protein of Newcastle disease virus localizes to the nucleus via a bipartite nuclear localization signal. Virology 195, 596-607.[CrossRef]
    [Google Scholar]
  5. Cormack, B. P., Valdivia, R. H. & Falkow, S. ( 1996; ). FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173, 33-38.[CrossRef]
    [Google Scholar]
  6. Dang, C. V. & Lee, W. M. F. ( 1989; ). Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. Journal of Biological Chemistry 264, 18019-18023.
    [Google Scholar]
  7. Davison, A. J. & Scott, J. E. ( 1986; ). The complete DNA sequence of varicella-zoster virus. Journal of General Virology 67, 1759-1816.[CrossRef]
    [Google Scholar]
  8. Dean, H. J. & Cheung, A. K. ( 1993; ). A 3′ coterminal gene cluster in pseudorabies virus contains herpes simplex virus UL1, UL2, and UL3 gene homologs and a unique UL3.5 open reading frame. Journal of Virology 67, 5955-5961.
    [Google Scholar]
  9. Desai, P. & Person, S. ( 1998; ). Incorporation of the green fluorescent protein into the herpes simplex virus type 1 capsid. Journal of Virology 72, 7563-7568.
    [Google Scholar]
  10. Dolan, A., Jamieson, F. E., Cunningham, C., Barnett, B. C. & McGeoch, D. J. ( 1998; ). The genome sequence of herpes simplex virus type 2. Journal of Virology 72, 2010-2021.
    [Google Scholar]
  11. Fankhauser, C., Izaurralde, E., Adachi, Y., Wingfield, P. & Laemmli, U. K. ( 1991; ). Specific complex of human immunodeficiency virus type 1 rev and nucleolar B23 proteins: dissociation by the Rev response element. Molecular and Cellular Biology 11, 2567-2575.
    [Google Scholar]
  12. Favre, D., Studer, E. & Michel, M. R. ( 1994; ). Two nucleolar targeting signals present in the N-terminal part of Semliki Forest virus capsid protein. Archives of Virology 137, 149-155.[CrossRef]
    [Google Scholar]
  13. Foster, T. P., Rybachuk, G. V. & Kousoulas, K. G. ( 1998; ). Expression of the enhanced green fluorescent protein by herpes simplex virus type 1 (HSV-1) as an in vitro or in vivo marker for virus entry and replication. Journal of Virological Methods 75, 151-160.[CrossRef]
    [Google Scholar]
  14. Georgopoulou, U., Kakkanas, A., Miriagou, V., Michaelidou, A. & Mavromara, P. ( 1995; ). Characterization of the US8.5 protein of herpes simplex virus. Archives of Virology 140, 2227-2241.[CrossRef]
    [Google Scholar]
  15. Ghiasi, H., Perng, G.-C., Cai, S., Nesburn, A. B. & Wechsler, S. L. ( 1996; ). The UL3 open reading frame of herpes simplex virus type 1 codes for a phosphoprotein. Virus Research 44, 137-142.[CrossRef]
    [Google Scholar]
  16. Harlow, E. & Lane, D. (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  17. Hauber, J., Malim, M. H. & Cullen, B. R. ( 1989; ). Mutational analysis of the conserved basic domain of human immunodeficiency virus tat protein. Journal of Virology 63, 1181-1187.
    [Google Scholar]
  18. Khattar, S. K., van Drunen Littel-van den Hurk, S., Babiuk, L. A. & Tikoo, S. K. ( 1995; ). Identification and transcriptional analysis of a 3′-coterminal gene cluster containing UL1, UL2, UL3, and UL3.5 open reading frames of bovine herpesvirus-1. Virology 213, 28-37.[CrossRef]
    [Google Scholar]
  19. Leopardi, R. & Roizman, B. ( 1996; ). Functional interaction and colocalization of the herpes simplex virus 1 major regulatory protein ICP4 with EAP, a nucleolar-ribosomal protein. Proceedings of the National Academy of Sciences, USA 93, 4572-4576.[CrossRef]
    [Google Scholar]
  20. Liu, J.-L., Lee, L. F., Ye, Y., Qian, Z. & Kung, H.-J. ( 1997; ). Nucleolar and nuclear localization properties of a herpesvirus bZIP oncoprotein, MEQ. Journal of Virology 71, 3188-3196.
    [Google Scholar]
  21. Lutz, P., Puvion-Dutilleul, F., Lutz, Y. & Kedinger, C. ( 1996; ). Nucleoplasmic and nucleolar distribution of the adenovirus IVa2 gene product. Journal of Virology 70, 3449-3460.
    [Google Scholar]
  22. McGeoch, D. J., Dalrymple, M. A., Davison, A. J., Dolan, A., Frame, M. C., McNab, D., Perry, L. J., Scott, J. E. & Taylor, P. ( 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.[CrossRef]
    [Google Scholar]
  23. McGeoch, D. J., Cunningham, C., McIntyre, G. & Dolan, A. ( 1991; ). Comparative sequence analysis of the long repeat regions and adjoining parts of the long unique regions in the genomes of herpes simplex viruses types 1 and 2. Journal of General Virology 72, 3057-3075.[CrossRef]
    [Google Scholar]
  24. MacLean, C. A., Rixon, F. J. & Marsden, H. S. ( 1987; ). The products of gene US11 of herpes simplex virus type 1 are DNA-binding and localize to the nucleoli of infected cells. Journal of General Virology 68, 1921-1937.[CrossRef]
    [Google Scholar]
  25. Mears, W. E., Lam, V. & Rice, S. A. ( 1995; ). Identification of nuclear and nucleolar localization signals in the herpes simplex virus regulatory protein ICP27. Journal of Virology 69, 935-947.
    [Google Scholar]
  26. Mèlëse, T. & Xue, Z. ( 1995; ). The nucleolus: an organelle formed by the act of building a ribosome. Current Opinion in Cell Biology 7, 319-324.[CrossRef]
    [Google Scholar]
  27. Miyazaki, Y., Takamatsu, T., Nosaka, T., Fujita, S., Martin, T. E. & Hatanaka, M. ( 1995; ). The cytotoxicity of human immunodeficiency virus type 1 Rev: implications for its interaction with the nucleolar protein B23. Experimental Cell Research 219, 93-101.[CrossRef]
    [Google Scholar]
  28. Peculis, B. A. & Gall, J. G. ( 1992; ). Localization of the nucleolar protein NO38 in amphibian oocytes. Journal of Cell Biology 116, 1-14.[CrossRef]
    [Google Scholar]
  29. Peränen, J., Rikkonen, M., Liljeström, P. & Kääriäinen, L. ( 1990; ). Nuclear localization of Semliki Forest virus-specific nonstructural protein nsP2. Journal of Virology 64, 1888-1896.
    [Google Scholar]
  30. Rikkonen, M., Peränen, J. & Kääriäinen, L. ( 1992; ). Nuclear and nucleolar targeting signals of Semliki Forest virus nonstructural protein nsP2. Virology 189, 462-473.[CrossRef]
    [Google Scholar]
  31. Roizman, B. & Sears, A. E. ( 1996; ). Herpes simplex viruses and their replication. In Fields Virology, pp. 2231-2295. Edited by B. N. Fields, D. M. Knipe & P. M. Howley. Philadelphia: Lippincott–Raven.
  32. Roller, R. J. & Roizman, B. ( 1992; ). The herpes simplex virus 1 RNA binding protein US11 is a virion component and associates with ribosomal 60S subunits. Journal of Virology 66, 3624-3632.
    [Google Scholar]
  33. Scheer, U. & Weisenberger, D. ( 1994; ). The nucleolus. Current Opinion in Cell Biology 6, 354-359.[CrossRef]
    [Google Scholar]
  34. Schmidt-Zachmann, M. S. & Nigg, E. A. ( 1993; ). Protein localization to the nucleolus: a search for targeting domains in nucleolin. Journal of Cell Science 105, 799-806.
    [Google Scholar]
  35. Siomi, H., Shida, H., Nam, S. H., Nosaka, T., Maki, M. & Hatanaka, M. ( 1988; ). Sequence requirements for nucleolar localization of human T cell leukemia virus type 1 pX protein, which regulates viral RNA processing. Cell 55, 197-209.[CrossRef]
    [Google Scholar]
  36. Szekely, L., Jiang, W.-Q., Pokrovskaja, K., Wiman, K. G., Klein, G. & Ringertz, N. ( 1995; ). Reversible nucleolar translocation of Epstein–Barr virus-encoded EBNA-5 and hsp70 proteins after exposure to heat shock or cell density congestion. Journal of General Virology 76, 2423-2432.[CrossRef]
    [Google Scholar]
  37. Telford, E. A. R., Watson, M. S., McBride, K. & Davison, A. J. ( 1992; ). The DNA sequence of equine herpesvirus-1. Virology 189, 304-316.[CrossRef]
    [Google Scholar]
  38. Telford, E. A. R., Watson, M. S., Perry, J., Cullinane, A. A. & Davison, A. J. ( 1998; ). The DNA sequence of equine herpesvirus-4. Journal of General Virology 79, 1197-1203.
    [Google Scholar]
  39. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350-4354.[CrossRef]
    [Google Scholar]
  40. Tsurumi, T., Maeno, K. & Nishiyama, Y. ( 1986; ). Molecular cloning of herpes simplex virus type 2 DNA. Journal of Biochemistry 99, 981-984.
    [Google Scholar]
  41. Ward, P. L., Ogle, W. O. & Roizman, B. ( 1996; ). Assemblons: nuclear structures defined by aggregation of immature capsids and some tegument proteins of herpes simplex virus 1. Journal of Virology 70, 4623-4631.
    [Google Scholar]
  42. Worrad, D. M. & Caradonna, S. ( 1993; ). The herpes simplex virus type 2 UL3 open reading frame encodes a nuclear localizing phosphoprotein. Virology 195, 364-376.[CrossRef]
    [Google Scholar]
  43. Yamada, H., Daikoku, T., Yamashita, Y., Jiang, Y.-M., Tsurumi, T. & Nishiyama, Y. ( 1997; ). The product of the US10 gene of herpes simplex virus type 1 is a capsid/tegument-associated phosphoprotein which copurifies with the nuclear matrix. Journal of General Virology 78, 2923-2931.
    [Google Scholar]
  44. Yamada, H., Jiang, Y.-M., Oshima, S.-i., Daikoku, T., Yamashita, Y., Tsurumi, T. & Nishiyama, Y. ( 1998; ). Characterization of the UL55 gene product of herpes simplex virus type 2. Journal of General Virology 79, 1989-1995.
    [Google Scholar]
  45. Yoshida, S., Lee, L. F., Yanagida, N. & Nazerian, K. ( 1994; ). Identification and characterization of a Marek’s disease virus gene homologous to glycoprotein L of herpes simplex virus. Virology 204, 414-419.[CrossRef]
    [Google Scholar]
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