This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Krosky, P. M.
Right arrow Articles by Sethna, P. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Krosky, P. M.
Right arrow Articles by Sethna, P. B.

 Previous Article  |  Next Article 

Journal of Virology, July 2003, p. 7720-7727, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.7720-7727.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Human Cytomegalovirus UL44 Protein Is a Substrate for the UL97 Protein Kinase

Paula M. Krosky,1,{dagger} Moon-Chang Baek,1,{ddagger} Wan Jin Jahng,1 Imma Barrera,2,§ Robert J. Harvey,2 Karen K. Biron,2 Donald M. Coen,1* and Phiroze B. Sethna2

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115,1 GlaxoSmithKline, Research Triangle Park, North Carolina 277092

Received 23 December 2002/ Accepted 16 April 2003

The human cytomegalovirus UL97 protein is an unusual protein kinase that is able to autophosphorylate and to phosphorylate certain exogenous substrates, including nucleoside analogs such as ganciclovir. However, no natural substrate of UL97 in infected cells has been identified. We report here that recombinant UL44 protein became radiolabeled when incubated with recombinant UL97 and [32P]ATP and that both proteins could be coimmunoprecipitated by an antibody that recognizes either protein. Subsequent studies showed that highly purified, recombinant UL97 phosphorylated purified, recombinant UL44. This phosphorylation occurred on serine and threonine residues and was sensitive to inhibition by maribavir and to a mutation that inactivates UL97 catalytic activity. Two-dimensional gel electrophoresis revealed the absence of specific phosphorylated forms of UL44 in immunoprecipitates from lysates of cells infected with a UL97 null mutant virus or with wild-type virus in the presence of maribavir. The results indicate that UL97 is sufficient to phosphorylate UL44 in vitro and is necessary for the normal phosphorylation of UL44 in infected cells. This strongly suggests that UL44 is a natural substrate of UL97.

* Corresponding author. Mailing address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Ave., Boston, MA 02115. Phone: (617) 432-1691. Fax: (617) 432-3833. E-mail: don_coen{at}hms.harvard.edu.

{dagger} Present address: Screening Technologies Branch, Developmental Therapeutics Program, NCI Frederick, Frederick, MD 21702.

{ddagger} Present address: Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.

§ Present address: Mount Sinai School of Medicine, New York, NY 10029-6574.

Journal of Virology, July 2003, p. 7720-7727, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.7720-7727.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wang, F.-Z., Roy, D., Gershburg, E., Whitehurst, C. B., Dittmer, D. P., Pagano, J. S. (2009). Maribavir Inhibits Epstein-Barr Virus Transcription in Addition to Viral DNA Replication. J. Virol. 83: 12108-12117 [Abstract] [Full Text]  
  • Chahine, M. N., Pierce, G. N. (2009). Therapeutic Targeting of Nuclear Protein Import in Pathological Cell Conditions. Pharmacol. Rev. 61: 358-372 [Abstract] [Full Text]  
  • Strang, B. L., Sinigalia, E., Silva, L. A., Coen, D. M., Loregian, A. (2009). Analysis of the Association of the Human Cytomegalovirus DNA Polymerase Subunit UL44 with the Viral DNA Replication Factor UL84. J. Virol. 83: 7581-7589 [Abstract] [Full Text]  
  • Thomas, M., Rechter, S., Milbradt, J., Auerochs, S., Muller, R., Stamminger, T., Marschall, M. (2009). Cytomegaloviral protein kinase pUL97 interacts with the nuclear mRNA export factor pUL69 to modulate its intranuclear localization and activity. J. Gen. Virol. 90: 567-578 [Abstract] [Full Text]  
  • Chou, S. (2009). Diverse Cytomegalovirus UL27 Mutations Adapt to Loss of Viral UL97 Kinase Activity under Maribavir. Antimicrob. Agents Chemother. 53: 81-85 [Abstract] [Full Text]  
  • Sinigalia, E., Alvisi, G., Mercorelli, B., Coen, D. M., Pari, G. S., Jans, D. A., Ripalti, A., Palu, G., Loregian, A. (2008). Role of Homodimerization of Human Cytomegalovirus DNA Polymerase Accessory Protein UL44 in Origin-Dependent DNA Replication in Cells. J. Virol. 82: 12574-12579 [Abstract] [Full Text]  
  • Kalejta, R. F. (2008). Tegument Proteins of Human Cytomegalovirus. Microbiol. Mol. Biol. Rev. 72: 249-265 [Abstract] [Full Text]  
  • Prichard, M. N., Sztul, E., Daily, S. L., Perry, A. L., Frederick, S. L., Gill, R. B., Hartline, C. B., Streblow, D. N., Varnum, S. M., Smith, R. D., Kern, E. R. (2008). Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes. J. Virol. 82: 5054-5067 [Abstract] [Full Text]  
  • Goldwater, D. R., Dougherty, C., Schumacher, M., Villano, S. A. (2008). Effect of Ketoconazole on the Pharmacokinetics of Maribavir in Healthy Adults. Antimicrob. Agents Chemother. 52: 1794-1798 [Abstract] [Full Text]  
  • Chou, S., Marousek, G. I. (2008). Accelerated Evolution of Maribavir Resistance in a Cytomegalovirus Exonuclease Domain II Mutant. J. Virol. 82: 246-253 [Abstract] [Full Text]  
  • Kamil, J. P., Coen, D. M. (2007). Human Cytomegalovirus Protein Kinase UL97 Forms a Complex with the Tegument Phosphoprotein pp65. J. Virol. 81: 10659-10668 [Abstract] [Full Text]  
  • Loregian, A., Sinigalia, E., Mercorelli, B., Palu, G., Coen, D. M. (2007). Binding parameters and thermodynamics of the interaction of the human cytomegalovirus DNA polymerase accessory protein, UL44, with DNA: implications for the processivity mechanism. Nucleic Acids Res 35: 4779-4791 [Abstract] [Full Text]  
  • Isomura, H., Stinski, M. F., Kudoh, A., Nakayama, S., Iwahori, S., Sato, Y., Tsurumi, T. (2007). The Late Promoter of the Human Cytomegalovirus Viral DNA Polymerase Processivity Factor Has an Impact on Delayed Early and Late Viral Gene Products but Not on Viral DNA Synthesis. J. Virol. 81: 6197-6206 [Abstract] [Full Text]  
  • Schregel, V., Auerochs, S., Jochmann, R., Maurer, K., Stamminger, T., Marschall, M. (2007). Mapping of a self-interaction domain of the cytomegalovirus protein kinase pUL97. J. Gen. Virol. 88: 395-404 [Abstract] [Full Text]  
  • Izumiya, Y., Izumiya, C., Van Geelen, A., Wang, D.-H., Lam, K. S., Luciw, P. A., Kung, H.-J. (2007). Kaposi's Sarcoma-Associated Herpesvirus-Encoded Protein Kinase and Its Interaction with K-bZIP. J. Virol. 81: 1072-1082 [Abstract] [Full Text]  
  • Asai, R., Kato, A., Kato, K., Kanamori-Koyama, M., Sugimoto, K., Sairenji, T., Nishiyama, Y., Kawaguchi, Y. (2006). Epstein-Barr Virus Protein Kinase BGLF4 Is a Virion Tegument Protein That Dissociates from Virions in a Phosphorylation-Dependent Process and Phosphorylates the Viral Immediate-Early Protein BZLF1.. J. Virol. 80: 5125-5134 [Abstract] [Full Text]  
  • Gershburg, E., Pagano, J. S. (2005). Epstein-Barr virus infections: prospects for treatment. J Antimicrob Chemother 56: 277-281 [Abstract] [Full Text]  
  • De Bolle, L., Naesens, L., De Clercq, E. (2005). Update on Human Herpesvirus 6 Biology, Clinical Features, and Therapy. Clin. Microbiol. Rev. 18: 217-245 [Abstract] [Full Text]  
  • Herget, T., Freitag, M., Morbitzer, M., Kupfer, R., Stamminger, T., Marschall, M. (2004). Novel Chemical Class of pUL97 Protein Kinase-Specific Inhibitors with Strong Anticytomegaloviral Activity. Antimicrob. Agents Chemother. 48: 4154-4162 [Abstract] [Full Text]  
  • Chou, S., Marousek, G. I., Senters, A. E., Davis, M. G., Biron, K. K. (2004). Mutations in the Human Cytomegalovirus UL27 Gene That Confer Resistance to Maribavir. J. Virol. 78: 7124-7130 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»