Browse > Article

Substrate Specificity of UL97 Protein Kinase from Human Cytomegalovirus using Spot Assay  

Baek, Moon-Chang (Department of Molecular Medicine, School of Medicine, Kyungpook National University)
Publication Information
YAKHAK HOEJI / v.50, no.4, 2006 , pp. 268-271 More about this Journal
Abstract
Protein kinase UL97 is an unusual protein kinase that can phosphorylate nucleoside analogs as well as protein/peptide. Previously we found a H2B-derived peptide, KESYSVYVYKV and reported that the P+5 position (K) is important. To further understand the substrate specificity at the P+5 position, we introduced spot assay system and showed that a peptide containing K residue among other amino acids at the P+5 position is the best substrate. Also other residues such as M, I, L, or G are good enough to be substrate of UL97. This result may aid the discovery of a new antiviral inhibitor.
Keywords
protein kinase UL97; peptide substrate; spot assay;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Krosky, P. M., Baek, M. C., Jahng, W. J., Barrera, I., Harvey, R. J., Biron, K. K., Coen, D. M. and Sethna, P. B. : The human cytomegalovirus UL44 protein is a substrate for the UL97 protein kinase. J. Virol. 77, 7720 (2003)   DOI   ScienceOn
2 Crossen, R. and Gruenwald, S. : In Baculovirus Expression Vector System Manual, 5th ed. Pharmingen, San Diego (1998)
3 He, Z., Kim, Y., Chu, L., Ohmstede, C., Biron, K. K. and Coen, D. M. : The human cytomegalovirus UL97 protein is a protein kinase that autophosphorylates on serines and threonines. J. Virol. 71, 405 (1997)
4 Selleseth, D. W., Talarico, C. L., Miller, T., Lutz, M. W., Biron, K. K. and Harvey, R. J. : In vitro activities of benzimidazole D- and L-ribonucleosides against herpesviruses. Antimicrob. Agents Chemother. 47, 2186 (2003)   DOI   ScienceOn
5 Talarico, C. L., Burnette, T. C., Miller, W. H., Smith, S. L., Davis, M. G., Stanat, S. C., Ng, T. I., He, Z., Coen, D. M., Roizman, B. and Biron, K. K. : Acyclovir is phosphorylated by the human cytomegalovirus UL97 protein. Antimicrob. Agents Chemother. 43, 1941 (1999)
6 Hanks, S. K. and Hunter, T. : Protein kinases 6. The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification. FASEB J. 9, 576 (1995)   DOI
7 Wolf, D. G., Courcelle, C. T., Prichard, M. N. and Mocarski, E. S. : Distinct and separate roles for herpesvirus-conserved UL97 kinase in cytomegalovirus DNA synthesis and encapsidation. Proc. Natl. Acad. Sci. U.S.A. 98, 1895 (2001)
8 Biron, K. K., Harvery, R. J., Chamberlain, S. C., Good, S. S., Smith, A. A. 3rd, Davis, M. G., Talarico, C. L., Miller, W. H., Ferris, R., Dornsife, R. E., Stanat, S. C., Drach, J. C., Townsed, L. B. and Koszalka, G. W. : Potent and selective inhibition of human cytomegalovirus replication by 1263W94, a benzimidazole L-riboside with a unique mode of action. Antimicrob. Agents Chemother. 46, 2365 (2002)   DOI   ScienceOn
9 Sullivan, V., Talarico, C. L., Stanat, S. C., Davis, M., Coen, D. M. and Biron, K. K. : A protein kinase homologue controls phosphorylation of ganciclovir in human cytomegalovirusinfected cells. Nature 358, 162 (1992)   DOI   ScienceOn
10 Littler, E., Stuart, A. D. and Chee, M. S. : Human cytomegalovirus UL97 open reading frame encodes a protein that phosphorylates the antiviral nucleoside analogue ganciclovir. Nature 358, 160 (1992)   DOI   ScienceOn
11 Baek, M. C., Krosky, P. M., He, Z. and Coen, D. M. : Specific phosphorylation of exogenous protein and peptide substrates by the human cytomegalovirus UL97 protein kinase. Importance of the P+5 position. J. Biol. Chem. 277, 29593 (2002)   DOI   ScienceOn
12 Krosky, P. M., Baek, M. C. and Coen, D. M. : The human cytomegalovirus UL97 protein kinase, an antiviral drug target, is required at the stage of nuclear egress. J. Virol. 77, 905 (2003)   DOI   ScienceOn
13 Baek, M. C., Krosky, P. M. and Coen, D. M. : Relationship between autophosphorylation and phosphorylation of exogenous substrates by the human cytomegalovirus UL97 protein kinase. J. Virol. 76, 11943 (2002)   DOI   ScienceOn
14 Songyang, Z., Lu, K. P., Kwon, Y. T., Tsai, L.-H., Filhol, O., Cochet, C., Brickey, D. A., Soderling, T. R., Bartleson, C., Graves, D. J., Demaggio, A. J., Hoekstra, M. F., Blenis, J., Hunter, T. and Cantley, L. C. : A structural basis for substrate specificities of protein Ser/Thr kinases: primary sequence preference of casein kinases I and II, NIMA, phosphorylase kinase, calmodulin-dependent kinase II, CDK5, and Erk. Mol. Cell. Biol. 16, 6486 (1996)   DOI
15 Prichard, M. N., Gao, N., Jairath, S., Mulamba, G., Krosky, P., Coen, D. M., Parker, B. O. and Pari, G. S. : A recombinant human cytomegalovirus with a large deletion in UL97 has a severe replication deficiency. J. Virol. 73, 5663 (1999)
16 Chee, M. S., Lawrence, G. L. and Barrell, B. G. : Alpha-, betaand gammaherpesviruses encode a putative phosphotransferase. J. Gen. Virol. 70, 1151 (1989)   DOI   ScienceOn