Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Jun 1;90(11):4991–4995. doi: 10.1073/pnas.90.11.4991

Maturation stage and proliferation-dependent expression of dUTPase in human T cells.

J R Strahler 1, X X Zhu 1, N Hora 1, Y K Wang 1, P C Andrews 1, N A Roseman 1, J V Neel 1, L Turka 1, S M Hanash 1
PMCID: PMC46639  PMID: 8389461

Abstract

We have developed a database of lymphoid polypeptides detected by two-dimensional polyacrylamide gel electrophoresis to aid in studies of leukemogenesis and of mutation affecting protein structure. In prior studies, we observed a 19-kDa phosphopolypeptide which was induced with proliferation in mature T cells and constitutively expressed in immature thymocytes. In this report we describe the identification of this polypeptide as the phosphorylated form of dUTPase (EC 3.6.1.23), following cDNA cloning of the gene, based on a partial amino acid sequence of the phosphopolypeptide. Studies of the expression and phosphorylation of dUTPase in human T cells indicate that accumulation and phosphorylation of dUTPase in mature T cells occur in a cell cycle-dependent manner. Interestingly, noncycling immature thymocytes express constitutively high levels of phosphorylated and unphosphorylated dUTPase. These results suggest an important role for dUTPase in immature thymocytes that is independent of proliferation.

Full text

PDF
4993

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen J. M., Forbush K. A., Perlmutter R. M. Functional dissection of the lck proximal promoter. Mol Cell Biol. 1992 Jun;12(6):2758–2768. doi: 10.1128/mcb.12.6.2758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bravo R., Frank R., Blundell P. A., Macdonald-Bravo H. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta. Nature. 1987 Apr 2;326(6112):515–517. doi: 10.1038/326515a0. [DOI] [PubMed] [Google Scholar]
  3. Brynolf K., Eliasson R., Reichard P. Formation of Okazaki fragments in polyoma DNA synthesis caused by misincorporation of uracil. Cell. 1978 Mar;13(3):573–580. doi: 10.1016/0092-8674(78)90330-6. [DOI] [PubMed] [Google Scholar]
  4. Crabtree G. R. Contingent genetic regulatory events in T lymphocyte activation. Science. 1989 Jan 20;243(4889):355–361. doi: 10.1126/science.2783497. [DOI] [PubMed] [Google Scholar]
  5. Goulian M., Bleile B. M., Dickey L. M., Grafstrom R. H., Ingraham H. A., Neynaber S. A., Peterson M. S., Tseng B. Y. Mechanism of thymineless death. Adv Exp Med Biol. 1986;195(Pt B):89–95. doi: 10.1007/978-1-4684-1248-2_15. [DOI] [PubMed] [Google Scholar]
  6. Grafstrom R. H., Tseng B. Y., Goulian M. The incorporation of uracil into animal cell DNA in vitro. Cell. 1978 Sep;15(1):131–140. doi: 10.1016/0092-8674(78)90089-2. [DOI] [PubMed] [Google Scholar]
  7. Gratiot-Deans J., Keim D., Strahler J. R., Turka L. A., Hanash S. Differential expression of Op18 phosphoprotein during human thymocyte maturation. J Clin Invest. 1992 Oct;90(4):1576–1581. doi: 10.1172/JCI116026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hanash S. M., Strahler J. R., Chan Y., Kuick R., Teichroew D., Neel J. V., Hailat N., Keim D. R., Gratiot-Deans J., Ungar D. Data base analysis of protein expression patterns during T-cell ontogeny and activation. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3314–3318. doi: 10.1073/pnas.90.8.3314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hanash S. M., Strahler J. R., Neel J. V., Hailat N., Melhem R., Keim D., Zhu X. X., Wagner D., Gage D. A., Watson J. T. Highly resolving two-dimensional gels for protein sequencing. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5709–5713. doi: 10.1073/pnas.88.13.5709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnston R. F., Pickett S. C., Barker D. L. Autoradiography using storage phosphor technology. Electrophoresis. 1990 May;11(5):355–360. doi: 10.1002/elps.1150110503. [DOI] [PubMed] [Google Scholar]
  11. Lindahl T. DNA glycosylases, endonucleases for apurinic/apyrimidinic sites, and base excision-repair. Prog Nucleic Acid Res Mol Biol. 1979;22:135–192. doi: 10.1016/s0079-6603(08)60800-4. [DOI] [PubMed] [Google Scholar]
  12. Lirette R., Caradonna S. Inhibition of phosphorylation of cellular dUTP nucleotidohydrolase as a consequence of herpes simplex virus infection. J Cell Biochem. 1990 Aug;43(4):339–353. doi: 10.1002/jcb.240430406. [DOI] [PubMed] [Google Scholar]
  13. Lundberg L. G., Thoresson H. O., Karlström O. H., Nyman P. O. Nucleotide sequence of the structural gene for dUTPase of Escherichia coli K-12. EMBO J. 1983;2(6):967–971. doi: 10.1002/j.1460-2075.1983.tb01529.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McIntosh E. M., Ager D. D., Gadsden M. H., Haynes R. H. Human dUTP pyrophosphatase: cDNA sequence and potential biological importance of the enzyme. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8020–8024. doi: 10.1073/pnas.89.17.8020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nation M. D., Guzder S. N., Giroir L. E., Deutsch W. A. Control of Drosophila deoxyuridine triphosphatase. Existence of a developmentally expressed protein inhibitor. Biochem J. 1989 Apr 15;259(2):593–596. doi: 10.1042/bj2590593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pardee A. B. G1 events and regulation of cell proliferation. Science. 1989 Nov 3;246(4930):603–608. doi: 10.1126/science.2683075. [DOI] [PubMed] [Google Scholar]
  17. Pardo E. G., Gutiérrez C. Cell cycle- and differentiation stage-dependent variation of dUTPase activity in higher plant cells. Exp Cell Res. 1990 Jan;186(1):90–98. doi: 10.1016/0014-4827(90)90214-u. [DOI] [PubMed] [Google Scholar]
  18. Prelich G., Stillman B. Coordinated leading and lagging strand synthesis during SV40 DNA replication in vitro requires PCNA. Cell. 1988 Apr 8;53(1):117–126. doi: 10.1016/0092-8674(88)90493-x. [DOI] [PubMed] [Google Scholar]
  19. Reinherz E. L., Schlossman S. F. The differentiation and function of human T lymphocytes. Cell. 1980 Apr;19(4):821–827. doi: 10.1016/0092-8674(80)90072-0. [DOI] [PubMed] [Google Scholar]
  20. Scharff M. D., Aguila H. L., Behar S. M., Chien N. C., DePinho R., French D. L., Pollock R. R., Shin S. U. Studies on the somatic instability of immunoglobulin genes in vivo and in cultured cells. Immunol Rev. 1987 Apr;96:75–90. doi: 10.1111/j.1600-065x.1987.tb00510.x. [DOI] [PubMed] [Google Scholar]
  21. Shivji K. K., Kenny M. K., Wood R. D. Proliferating cell nuclear antigen is required for DNA excision repair. Cell. 1992 Apr 17;69(2):367–374. doi: 10.1016/0092-8674(92)90416-a. [DOI] [PubMed] [Google Scholar]
  22. Slabaugh M. B., Roseman N. A. Retroviral protease-like gene in the vaccinia virus genome. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4152–4155. doi: 10.1073/pnas.86.11.4152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Spindler K. R., Rosser D. S., Berk A. J. Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. J Virol. 1984 Jan;49(1):132–141. doi: 10.1128/jvi.49.1.132-141.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Strominger J. L. Developmental biology of T cell receptors. Science. 1989 May 26;244(4907):943–950. doi: 10.1126/science.2658058. [DOI] [PubMed] [Google Scholar]
  25. Takahama Y., Singer A. Post-transcriptional regulation of early T cell development by T cell receptor signals. Science. 1992 Nov 27;258(5087):1456–1462. doi: 10.1126/science.1439838. [DOI] [PubMed] [Google Scholar]
  26. Thompson C. B., Challoner P. B., Neiman P. E., Groudine M. Expression of the c-myb proto-oncogene during cellular proliferation. 1986 Jan 30-Feb 5Nature. 319(6052):374–380. doi: 10.1038/319374a0. [DOI] [PubMed] [Google Scholar]
  27. Turka L. A., Ledbetter J. A., Lee K., June C. H., Thompson C. B. CD28 is an inducible T cell surface antigen that transduces a proliferative signal in CD3+ mature thymocytes. J Immunol. 1990 Mar 1;144(5):1646–1653. [PubMed] [Google Scholar]
  28. Turka L. A., Linsley P. S., Paine R., 3rd, Schieven G. L., Thompson G. B., Ledbetter J. A. Signal transduction via CD4, CD8, and CD28 in mature and immature thymocytes. Implications for thymic selection. J Immunol. 1991 Mar 1;146(5):1428–1436. [PubMed] [Google Scholar]
  29. Tye B. K., Lehman I. R. Excision repair of uracil incorporated in DNA as a result of a defect in dUTPase. J Mol Biol. 1977 Dec 5;117(2):293–306. doi: 10.1016/0022-2836(77)90128-0. [DOI] [PubMed] [Google Scholar]
  30. Wildin R. S., Garvin A. M., Pawar S., Lewis D. B., Abraham K. M., Forbush K. A., Ziegler S. F., Allen J. M., Perlmutter R. M. Developmental regulation of lck gene expression in T lymphocytes. J Exp Med. 1991 Feb 1;173(2):383–393. doi: 10.1084/jem.173.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zhu X. X., Kozarsky K., Strahler J. R., Eckerskorn C., Lottspeich F., Melhem R., Lowe J., Fox D. A., Hanash S. M., Atweh G. F. Molecular cloning of a novel human leukemia-associated gene. Evidence of conservation in animal species. J Biol Chem. 1989 Aug 25;264(24):14556–14560. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES