Abstract
The serine/threonine kinase encoded by the Us3 gene is conserved amongst all known alphaherpesviruses. Us3 has been reported to function in a variety of aspects of the virus lifecycle including protection of cells from virus-induced apoptosis, de-envelopment of enveloped virus particles from the perinuclear space and cell-to-cell spread of virus infection. In this report, we examined the sub-cellular localization of the pseudorabies virus (PRV) Us3 homolog. The PRV Us3 gene encodes two proteins termed Us3a and Us3b. Us3a differs from Us3b in that it contains 54 additional N-terminal amino acids. In transfected cells, Us3a localized predominantly to the plasma membrane whereas the Us3b protein localized predominantly to the nucleus. To explore the differences in the localization of the Us3a and Us3b proteins, we fused the amino-terminal 54 amino acids of Us3a to the amino-terminus of the enhanced green fluorescent protein (EGFP). Surprisingly, this fusion protein localized exclusively to mitochondria in transfected cells. Analysis of mutated Us3–EGFP fusion proteins in transfected cells revealed that the carboxy-terminal 101 amino acids of Us3a and Us3b comprises a membrane/vesicular localization domain, and that the N-terminal 102 amino acids of Us3b comprises a nuclear localization domain. We provide a model to rationalize the complex localization of Us3a and Us3b in transfected cells and hypothesize that the mitochondrial, nuclear and membrane localization motifs function in the reported anti-apoptotic, egress and cell-to-cell spread functions of Us3.
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Frame M.C., Purves F.C., McGeoch D.J., Marsden H.S., and Leader D.P. J Gen Virol 68, 2699–2704, 1987.
McGeoch D.J. and Davison A.J., Nucl Acids Res 14, 1765–1777, 1986.
Heineman T.C., Seidel K., and Cohen J.I., J Virol 70, 7312–7317, 1996.
Leopardi R., Van Sant C., and Roizman B., Proc Natl Acad Sci USA 94, 7891–7896, 1997.
Takashima Y., Tamura H., Xuan X., and Otsuka H., Virus Res 59, 23–34, 1999.
Wagenaar F., Pol J.M., Peeters B., Gielkens A.L., de Wind N., and Kimman T.G., J Gen Virol 76, 1851–1859, 1995.
Klupp B.G., Granzow H., and Mettenleiter T.C., J Gen Virol 82, 2363–2371, 2001.
Demmin G.L., Clase A.C., Randall J.A., Enquist L.W., and Banfield B.W., J Virol 75, 10856–10869, 2001.
Mettenleiter T.C., Vet Res 31, 99–115, 2000.
Rixon F.J. and McGeoch D.J., Nucl Acids Res 13, 953–973, 1985.
van Zijl M., van der Gulden H., de Wind N., Gielkens A., and Berns A., J Gen Virol 71, 1747–1755, 1990.
Zhang G. Stevens R., and Leader D.P., J Gen Virol 71, 1757–1765, 1990.
Lyman M.G., Demmin G.L., and Banfield B.W., J Virol 77, 1403–1414, 2003.
Schatz G., J Biol Chem 271, 31763–31766, 1996.
Kimman T.G., de Wind N., Oei-Lie N., Pol J. M., Berns A.J., and Gielkens A.L., J Gen Virol 73, 243–251, 1992.
Kimman T.G., De Wind N., De Bruin T., de Visser Y., and Voermans, J., Virology 205, 511–518, 1994.
Longnecker R. and Roizman B., Science 236, 573–576, 1987.
Nishiyama Y., Yamada Y., Kurachi R., and Daikoku T., Virology 190, 256–268, 1992.
Meignier B., Longnecker R., Mavromara-Nazos P., Sears A. E., and Roizman B., Virology 162, 251–254, 1988.
Corbett A.H. and Silver P.A., Microbiol Mol Biol Rev 61, 193–211, 1997.
Macara I.G., Microbiol Mol Biol Rev 65, 570–594, 2001.
van Leeuwen H., Elliott G., and O'Hare P., J Virol 76, 3471–3481, 2002.
van Minnebruggen G., Favoreel H.W., Jacobs L., and Nauwynck H.J., J Virol 77, 9074–9080, 2003.
Gavel Y. and von Heijne G., Protein Eng 4, 33–37, 1990.
Herrmann J.M. and Neupert W., Curr Opin Microbiol 3, 210–214, 2000.
Munger J., Chee, A.V., and Roizman, B., J Virol 75, 5491–5497, 2001.
Munger J. and Roizman B. Proc Natl Acad Sci USA 98, 10410–10415, 2001.
Jacotot E., Ferri K.F., El Hamel C., Brenner C., Druillennec S., Hoebeke J., Rustin P., Metivier D., Lenoir C., Geuskens M., Vieira H.L., Loeffler M., Belzacq A.S., Briand, J.P., Zamzami N., Edelman L., Xie Z.H., Reed J.C., Roques B.P., and Kroemer G., J Exp Med 193, 509–519, 2001.
Jacotot E., Ravagnan L., Loeffler M., Ferri K.F., Vieira H.L., Zamzami N., Costantini P., Druillennec S., Hoebeke J., Briand J.P., Irinopoulou T., Daugas E., Susin S.A., Cointe D., Xie Z.H., Reed J.C., Roques B.P., and Kroemer G., J Exp Med 191, 33–46, 2000.
Ciminale V., Zotti L., D'Agostino D.M., Ferro T., Casareto L., Franchini G., Bernardi P., and Chieco-Bianchi L., Oncogene 18, 4505–4514, 1999.
Goldmacher V.S., Bartle L.M., Skaletskaya A., Dionne C.A., Kedersha N.L., Vater C.A., Han J.W., Lutz R.J., Watanabe S., Cahir McFarland E.D., Kieff E.D., Mocarski E.S., and Chittenden T., Proc Natl Acad Sci USA 96, 12536–12541, 1999.
Takada S., Shirakata Y., Kaneniwa N., and Koike K., Oncogene 18, 6965–6973, 1999.
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Calton, C.M., Randall, J.A., Adkins, M.W. et al. The Pseudorabies Virus Serine/Threonine Kinase Us3 Contains Mitochondrial, Nuclear and Membrane Localization Signals. Virus Genes 29, 131–145 (2004). https://doi.org/10.1023/B:VIRU.0000032796.27878.7f
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DOI: https://doi.org/10.1023/B:VIRU.0000032796.27878.7f