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. 1993 Feb;13(2):1202–1211. doi: 10.1128/mcb.13.2.1202

Control of simian virus 40 DNA replication by the HeLa cell nuclear kinase casein kinase I.

A Cegielska 1, D M Virshup 1
PMCID: PMC359005  PMID: 8380893

Abstract

The initiation of simian virus 40 (SV40) DNA replication is regulated by the phosphorylation state of the viral initiator protein, large T antigen. We describe the purification from HeLa cell nuclei of a 35-kDa serine/threonine protein kinase that phosphorylates T antigen at sites that are phosphorylated in vivo and thereby inhibits its ability to initiate SV40 DNA replication. The inhibition of both origin unwinding and DNA replication by the kinase is reversed by protein phosphatase 2A. As determined by molecular weight, substrate specificity, autophosphorylation, immunoreactivity, and limited sequence analysis, this kinase appears to be identical to casein kinase I, a ubiquitous serine/threonine protein kinase that is closely related to a yeast kinase involved in DNA metabolism. The HeLa cell phosphorylation cycle that controls the initiation of SV40 DNA replication may also play a role in cellular DNA metabolism.

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Selected References

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  1. Aebersold R. H., Leavitt J., Saavedra R. A., Hood L. E., Kent S. B. Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6970–6974. doi: 10.1073/pnas.84.20.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Agostinis P., Pinna L. A., Meggio F., Marin O., Goris J., Vandenheede J. R., Merlevede W. A synthetic peptide substrate specific for casein kinase I. FEBS Lett. 1989 Dec 18;259(1):75–78. doi: 10.1016/0014-5793(89)81498-x. [DOI] [PubMed] [Google Scholar]
  3. Borowiec J. A., Dean F. B., Bullock P. A., Hurwitz J. Binding and unwinding--how T antigen engages the SV40 origin of DNA replication. Cell. 1990 Jan 26;60(2):181–184. doi: 10.1016/0092-8674(90)90730-3. [DOI] [PubMed] [Google Scholar]
  4. Brockman J. L., Anderson R. A. Casein kinase I is regulated by phosphatidylinositol 4,5-bisphosphate in native membranes. J Biol Chem. 1991 Feb 5;266(4):2508–2512. [PubMed] [Google Scholar]
  5. Challberg M. D., Kelly T. J. Animal virus DNA replication. Annu Rev Biochem. 1989;58:671–717. doi: 10.1146/annurev.bi.58.070189.003323. [DOI] [PubMed] [Google Scholar]
  6. Chen Y. R., Lees-Miller S. P., Tegtmeyer P., Anderson C. W. The human DNA-activated protein kinase phosphorylates simian virus 40 T antigen at amino- and carboxy-terminal sites. J Virol. 1991 Oct;65(10):5131–5140. doi: 10.1128/jvi.65.10.5131-5140.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clertant P., Cuzin F. Covalent affinity labeling by periodate-oxidized [alpha-32P]ATP of the large-T proteins of polyoma and SV40 viruses. J Biol Chem. 1982 Jun 10;257(11):6300–6305. [PubMed] [Google Scholar]
  8. D'Urso G., Marraccino R. L., Marshak D. R., Roberts J. M. Cell cycle control of DNA replication by a homologue from human cells of the p34cdc2 protein kinase. Science. 1990 Nov 9;250(4982):786–791. doi: 10.1126/science.2173140. [DOI] [PubMed] [Google Scholar]
  9. DeGuzman A., Lee E. Y. Preparation of low-molecular-weight forms of rabbit muscle protein phosphatase. Methods Enzymol. 1988;159:356–368. doi: 10.1016/0076-6879(88)59036-5. [DOI] [PubMed] [Google Scholar]
  10. Dean F. B., Bullock P., Murakami Y., Wobbe C. R., Weissbach L., Hurwitz J. Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication. Proc Natl Acad Sci U S A. 1987 Jan;84(1):16–20. doi: 10.1073/pnas.84.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dutta A., Stillman B. cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication. EMBO J. 1992 Jun;11(6):2189–2199. doi: 10.1002/j.1460-2075.1992.tb05278.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fanning E., Knippers R. Structure and function of simian virus 40 large tumor antigen. Annu Rev Biochem. 1992;61:55–85. doi: 10.1146/annurev.bi.61.070192.000415. [DOI] [PubMed] [Google Scholar]
  14. Flotow H., Graves P. R., Wang A. Q., Fiol C. J., Roeske R. W., Roach P. J. Phosphate groups as substrate determinants for casein kinase I action. J Biol Chem. 1990 Aug 25;265(24):14264–14269. [PubMed] [Google Scholar]
  15. Flotow H., Roach P. J. Role of acidic residues as substrate determinants for casein kinase I. J Biol Chem. 1991 Feb 25;266(6):3724–3727. [PubMed] [Google Scholar]
  16. Flotow H., Roach P. J. Synergistic phosphorylation of rabbit muscle glycogen synthase by cyclic AMP-dependent protein kinase and casein kinase I. Implications for hormonal regulation of glycogen synthase. J Biol Chem. 1989 Jun 5;264(16):9126–9128. [PubMed] [Google Scholar]
  17. Grässer F. A., Scheidtmann K. H., Tuazon P. T., Traugh J. A., Walter G. In vitro phosphorylation of SV40 large T antigen. Virology. 1988 Jul;165(1):13–22. doi: 10.1016/0042-6822(88)90653-8. [DOI] [PubMed] [Google Scholar]
  18. Hoekstra M. F., Liskay R. M., Ou A. C., DeMaggio A. J., Burbee D. G., Heffron F. HRR25, a putative protein kinase from budding yeast: association with repair of damaged DNA. Science. 1991 Aug 30;253(5023):1031–1034. doi: 10.1126/science.1887218. [DOI] [PubMed] [Google Scholar]
  19. Hurwitz J., Dean F. B., Kwong A. D., Lee S. H. The in vitro replication of DNA containing the SV40 origin. J Biol Chem. 1990 Oct 25;265(30):18043–18046. [PubMed] [Google Scholar]
  20. Höss A., Moarefi I., Scheidtmann K. H., Cisek L. J., Corden J. L., Dornreiter I., Arthur A. K., Fanning E. Altered phosphorylation pattern of simian virus 40 T antigen expressed in insect cells by using a baculovirus vector. J Virol. 1990 Oct;64(10):4799–4807. doi: 10.1128/jvi.64.10.4799-4807.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lees-Miller S. P., Chen Y. R., Anderson C. W. Human cells contain a DNA-activated protein kinase that phosphorylates simian virus 40 T antigen, mouse p53, and the human Ku autoantigen. Mol Cell Biol. 1990 Dec;10(12):6472–6481. doi: 10.1128/mcb.10.12.6472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mastrangelo I. A., Hough P. V., Wall J. S., Dodson M., Dean F. B., Hurwitz J. ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication. Nature. 1989 Apr 20;338(6217):658–662. doi: 10.1038/338658a0. [DOI] [PubMed] [Google Scholar]
  23. McVey D., Brizuela L., Mohr I., Marshak D. R., Gluzman Y., Beach D. Phosphorylation of large tumour antigen by cdc2 stimulates SV40 DNA replication. Nature. 1989 Oct 12;341(6242):503–507. doi: 10.1038/341503a0. [DOI] [PubMed] [Google Scholar]
  24. O'Reilly D. R., Miller L. K. Expression and complex formation of simian virus 40 large T antigen and mouse p53 in insect cells. J Virol. 1988 Sep;62(9):3109–3119. doi: 10.1128/jvi.62.9.3109-3119.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pages J., Manteuil S., Stehelin D., Fiszman M., Marx M., Girard M. Relationship between replication of simian virus 40 DNA and specific events of the host cell cycle. J Virol. 1973 Jul;12(1):99–107. doi: 10.1128/jvi.12.1.99-107.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Parsons R. E., Stenger J. E., Ray S., Welker R., Anderson M. E., Tegtmeyer P. Cooperative assembly of simian virus 40 T-antigen hexamers on functional halves of the replication origin. J Virol. 1991 Jun;65(6):2798–2806. doi: 10.1128/jvi.65.6.2798-2806.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Prives C. The replication functions of SV40 T antigen are regulated by phosphorylation. Cell. 1990 Jun 1;61(5):735–738. doi: 10.1016/0092-8674(90)90179-i. [DOI] [PubMed] [Google Scholar]
  28. Roberts J. M., D'Urso G. An origin unwinding activity regulates initiation of DNA replication during mammalian cell cycle. Science. 1988 Sep 16;241(4872):1486–1489. doi: 10.1126/science.2843984. [DOI] [PubMed] [Google Scholar]
  29. Robinson L. C., Hubbard E. J., Graves P. R., DePaoli-Roach A. A., Roach P. J., Kung C., Haas D. W., Hagedorn C. H., Goebl M., Culbertson M. R. Yeast casein kinase I homologues: an essential gene pair. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):28–32. doi: 10.1073/pnas.89.1.28. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rowles J., Slaughter C., Moomaw C., Hsu J., Cobb M. H. Purification of casein kinase I and isolation of cDNAs encoding multiple casein kinase I-like enzymes. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9548–9552. doi: 10.1073/pnas.88.21.9548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ruediger R., Van Wart Hood J. E., Mumby M., Walter G. Constant expression and activity of protein phosphatase 2A in synchronized cells. Mol Cell Biol. 1991 Aug;11(8):4282–4285. doi: 10.1128/mcb.11.8.4282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Scheidtmann K. H., Buck M., Schneider J., Kalderon D., Fanning E., Smith A. E. Biochemical characterization of phosphorylation site mutants of simian virus 40 large T antigen: evidence for interaction between amino- and carboxy-terminal domains. J Virol. 1991 Mar;65(3):1479–1490. doi: 10.1128/jvi.65.3.1479-1490.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Scheidtmann K. H., Echle B., Walter G. Simian virus 40 large T antigen is phosphorylated at multiple sites clustered in two separate regions. J Virol. 1982 Oct;44(1):116–133. doi: 10.1128/jvi.44.1.116-133.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Scheidtmann K. H. Phosphorylation of simian virus 40 large T antigen: cytoplasmic and nuclear phophorylation sites differ in their metabolic stability. Virology. 1986 Apr 15;150(1):85–95. [PubMed] [Google Scholar]
  35. Scheidtmann K. H., Virshup D. M., Kelly T. J. Protein phosphatase 2A dephosphorylates simian virus 40 large T antigen specifically at residues involved in regulation of DNA-binding activity. J Virol. 1991 Apr;65(4):2098–2101. doi: 10.1128/jvi.65.4.2098-2101.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schneider J., Fanning E. Mutations in the phosphorylation sites of simian virus 40 (SV40) T antigen alter its origin DNA-binding specificity for sites I or II and affect SV40 DNA replication activity. J Virol. 1988 May;62(5):1598–1605. doi: 10.1128/jvi.62.5.1598-1605.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stahl H., Dröge P., Knippers R. DNA helicase activity of SV40 large tumor antigen. EMBO J. 1986 Aug;5(8):1939–1944. doi: 10.1002/j.1460-2075.1986.tb04447.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stillman B. Initiation of eukaryotic DNA replication in vitro. Annu Rev Cell Biol. 1989;5:197–245. doi: 10.1146/annurev.cb.05.110189.001213. [DOI] [PubMed] [Google Scholar]
  39. Tuazon P. T., Traugh J. A. Casein kinase I and II--multipotential serine protein kinases: structure, function, and regulation. Adv Second Messenger Phosphoprotein Res. 1991;23:123–164. [PubMed] [Google Scholar]
  40. Umphress J. L., Tuazon P. T., Chen C. J., Traugh J. A. Determinants on simian virus 40 large T antigen are important for recognition and phosphorylation by casein kinase I. Eur J Biochem. 1992 Jan 15;203(1-2):239–243. doi: 10.1111/j.1432-1033.1992.tb19852.x. [DOI] [PubMed] [Google Scholar]
  41. Virshup D. M., Kauffman M. G., Kelly T. J. Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A. EMBO J. 1989 Dec 1;8(12):3891–3898. doi: 10.1002/j.1460-2075.1989.tb08568.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Virshup D. M., Kelly T. J. Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro. Proc Natl Acad Sci U S A. 1989 May;86(10):3584–3588. doi: 10.1073/pnas.86.10.3584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Virshup D. M., Russo A. A., Kelly T. J. Mechanism of activation of simian virus 40 DNA replication by protein phosphatase 2A. Mol Cell Biol. 1992 Nov;12(11):4883–4895. doi: 10.1128/mcb.12.11.4883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wessel R., Schweizer J., Stahl H. Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication. J Virol. 1992 Feb;66(2):804–815. doi: 10.1128/jvi.66.2.804-815.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wold M. S., Kelly T. Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2523–2527. doi: 10.1073/pnas.85.8.2523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wold M. S., Weinberg D. H., Virshup D. M., Li J. J., Kelly T. J. Identification of cellular proteins required for simian virus 40 DNA replication. J Biol Chem. 1989 Feb 15;264(5):2801–2809. [PubMed] [Google Scholar]

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