Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Nov 1;107(5):1629–1642. doi: 10.1083/jcb.107.5.1629

The nuclear matrix protein, numatrin (B23), is associated with growth factor-induced mitogenesis in Swiss 3T3 fibroblasts and with T lymphocyte proliferation stimulated by lectins and anti-T cell antigen receptor antibody

PMCID: PMC2115313  PMID: 3141428

Abstract

Numatrin is a tightly bound nuclear matrix protein (40 kD/pI-5) whose synthesis is markedly and promptly increased in association with cellular commitment for mitogenesis in B lymphocytes. (Feuerstein, N., and J.J. Mond. 1987. J. Biol. Chem. 262:11389-11397). To study whether this event is exclusively associated with proliferation of B lymphocytes, we examined the synthesis of numatrin in T lymphocytes (murine and human) activated by lectins or by anti-T cell antigen receptor monoclonal antibody and in Swiss 3T3 fibroblasts stimulated by growth factors. We showed a close correlation between induction of DNA synthesis and induction of numatrin synthesis in T lymphocytes stimulated by concanavalin A, anti-T cell antigen receptor monoclonal antibody, and IL-2 in murine T cells. Similar results were observed in Swiss 3T3 fibroblasts, thus only combinations of growth factors (insulin/EGF or insulin/B subunit of cholera toxin) or serum, which induced a significant increase in DNA synthesis, were also associated with a significant increase in synthesis of numatrin. Similar to B cells, the increase in numatrin synthesis in fibroblasts was found to occur at early G1 phase. The calcium ionophores, A23187 and ionomycin, previously shown to induce an increase in c-myc and c-fos mRNA levels in fibroblasts, induced a marked increase in the synthesis of a nuclear protein at 80 kD/pI-5 but failed to induce an increase in the synthesis of numatrin indicating that an increase in intracellular Ca++ level is not sufficient for induction of the synthesis of numatrin. This further indicates that the increase in synthesis of numatrin may be more closely correlated with cellular commitment for mitogenesis as compared with other biochemical parameters. Using a polyclonal numatrin antibody we demonstrated that mitogen stimulation is also associated with a marked increase in numatrin abundance, which reached a peak at the onset of S phase and declined at the end of S phase. Evidence is presented to show that numatrin synthesis and abundance is elevated in various lymphoma cell lines. Using indirect immunofluorescence assays we showed that numatrin is abundant in other malignant cells: KB, epidermoid carcinoma, and Hep2 human hepatoma. Immunofluorescence studies further showed that mitogen stimulation of B lymphocytes induced a marked accumulation of numatrin in the nucleoli. This observation is in accord with the recent finding of identity of numatrin with the nucleolar protein B23 (Feuerstein et al. 1988. J. Biol. Chem. 263:10608-10612).(ABSTRACT TRUNCATED AT 400 WORDS)

Full Text

The Full Text of this article is available as a PDF (4.2 MB).

Selected References

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

  1. Ballal N. R., Goldknopf I. L., Goldberg D. A., Busch H. The dynamic state of liver nucleolar proteins as reflected by their changes during administration of thioacetamide. Life Sci. 1974 May 16;14(10):1835–1845. doi: 10.1016/0024-3205(74)90401-9. [DOI] [PubMed] [Google Scholar]
  2. Bishop J. M. Viral oncogenes. Cell. 1985 Aug;42(1):23–38. doi: 10.1016/s0092-8674(85)80098-2. [DOI] [PubMed] [Google Scholar]
  3. Bodnar J. W., Jones C. J., Coombs D. H., Pearson G. D., Ward D. C. Proteins tightly bound to HeLa cell DNA at nuclear matrix attachment sites. Mol Cell Biol. 1983 Sep;3(9):1567–1579. doi: 10.1128/mcb.3.9.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braverman R., Bhattacharya B., Feuerstein N., Cooper H. L. Identification and characterization of the nonphosphorylated precursor of pp17, a phosphoprotein associated with phorbol ester induction of growth arrest and monocytic differentiation in HL-60 promyelocytic leukemia cells. J Biol Chem. 1986 Oct 25;261(30):14342–14348. [PubMed] [Google Scholar]
  5. Busch R. K., Chan P. K., Busch H. Actinomycin D inhibition of monoclonal antibody binding to nucleolar phosphoprotein 37/5.2 (B23). Life Sci. 1984 Oct 22;35(17):1777–1785. doi: 10.1016/0024-3205(84)90275-3. [DOI] [PubMed] [Google Scholar]
  6. Celis J. E., Celis A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Proc Natl Acad Sci U S A. 1985 May;82(10):3262–3266. doi: 10.1073/pnas.82.10.3262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Celis J. E., Nielsen S. Proliferation-sensitive nuclear phosphoprotein "dividin" is synthesized almost exclusively during S phase of the cell cycle in human AMA cells. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8187–8190. doi: 10.1073/pnas.83.21.8187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chan P. K., Aldrich M., Busch H. Alterations in immunolocalization of the phosphoprotein B23 in HeLa cells during serum starvation. Exp Cell Res. 1985 Nov;161(1):101–110. doi: 10.1016/0014-4827(85)90494-x. [DOI] [PubMed] [Google Scholar]
  9. Dicker P., Rozengurt E. Phorbol esters and vasopressin stimulate DNA synthesis by a common mechanism. Nature. 1980 Oct 16;287(5783):607–612. doi: 10.1038/287607a0. [DOI] [PubMed] [Google Scholar]
  10. Feuerstein N., Ali I. U. Comparative analysis of p21 proteins from various cell types by two-dimensional gel electrophoresis. J Cell Biochem. 1985;29(3):253–263. doi: 10.1002/jcb.240290309. [DOI] [PubMed] [Google Scholar]
  11. Feuerstein N., Chan P. K., Mond J. J. Identification of numatrin, the nuclear matrix protein associated with induction of mitogenesis, as the nucleolar protein B23. Implication for the role of the nucleolus in early transduction of mitogenic signals. J Biol Chem. 1988 Aug 5;263(22):10608–10612. [PubMed] [Google Scholar]
  12. Feuerstein N., Cooper H. L. Rapid protein phosphorylation induced by phorbol ester in HL-60 cells. Unique alkali-stable phosphorylation of a 17,000-dalton protein detected by two-dimensional gel electrophoresis. J Biol Chem. 1983 Sep 10;258(17):10786–10793. [PubMed] [Google Scholar]
  13. Feuerstein N., Mond J. J. "Numatrin," a nuclear matrix protein associated with induction of proliferation in B lymphocytes. J Biol Chem. 1987 Aug 15;262(23):11389–11397. [PubMed] [Google Scholar]
  14. Feuerstein N., Mond J. J. Identification of a prominent nuclear protein associated with proliferation of normal and malignant B cells. J Immunol. 1987 Sep 15;139(6):1818–1822. [PubMed] [Google Scholar]
  15. Kang Y. J., Olson M. O., Busch H. Phosphorylation of acid-soluble proteins in isolated nucleoli of Novikoff hepatoma ascites cells. Effects of divalent cations. J Biol Chem. 1974 Sep 10;249(17):5580–5585. [PubMed] [Google Scholar]
  16. Kang Y. J., Olson M. O., Jones C., Busch H. Nucleolar phosphoproteins of normal rat liver and Novikoff hepatoma ascites cells. Cancer Res. 1975 Jun;35(6):1470–1475. [PubMed] [Google Scholar]
  17. Mond J. J., Mongini P. K., Sieckmann D., Paul W. E. Role of T lymphocytes in the response to TNP-AECM-Ficoll. J Immunol. 1980 Sep;125(3):1066–1070. [PubMed] [Google Scholar]
  18. Ran W., Dean M., Levine R. A., Henkle C., Campisi J. Induction of c-fos and c-myc mRNA by epidermal growth factor or calcium ionophore is cAMP dependent. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8216–8220. doi: 10.1073/pnas.83.21.8216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reich N. C., Levine A. J. Growth regulation of a cellular tumour antigen, p53, in nontransformed cells. Nature. 1984 Mar 8;308(5955):199–201. doi: 10.1038/308199a0. [DOI] [PubMed] [Google Scholar]
  20. Rozengurt E. Early signals in the mitogenic response. Science. 1986 Oct 10;234(4773):161–166. doi: 10.1126/science.3018928. [DOI] [PubMed] [Google Scholar]
  21. Spiegel S., Fishman P. H. Gangliosides as bimodal regulators of cell growth. Proc Natl Acad Sci U S A. 1987 Jan;84(1):141–145. doi: 10.1073/pnas.84.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tsokos G. C., Magrath I. T., Balow J. E. Epstein-Barr virus induces normal B cell responses but defective suppressor T cell responses in patients with systemic lupus erythematosus. J Immunol. 1983 Oct;131(4):1797–1801. [PubMed] [Google Scholar]
  23. Tsuda T., Hamamori Y., Yamashita T., Fukumoto Y., Takai Y. Involvement of three intracellular messenger systems, protein kinase C, calcium ion and cyclic AMP, in the regulation of c-fos gene expression in Swiss 3T3 cells. FEBS Lett. 1986 Nov 10;208(1):39–42. doi: 10.1016/0014-5793(86)81527-7. [DOI] [PubMed] [Google Scholar]
  24. Tsuda T., Kaibuchi K., West B., Takai Y. Involvement of Ca2+ in platelet-derived growth factor-induced expression of c-myc oncogene in Swiss 3T3 fibroblasts. FEBS Lett. 1985 Jul 22;187(1):43–46. doi: 10.1016/0014-5793(85)81210-2. [DOI] [PubMed] [Google Scholar]
  25. Yamashita T., Tsuda T., Hamamori Y., Takai Y. Possible involvement of cyclic AMP and calcium ion in prostaglandin E1-induced elevation of c-myc mRNA levels in Swiss 3T3 fibroblasts. J Biol Chem. 1986 Dec 25;261(36):16878–16882. [PubMed] [Google Scholar]
  26. Yung B. Y., Busch H., Chan P. K. Translocation of nucleolar phosphoprotein B23 (37 kDa/pI 5.1) induced by selective inhibitors of ribosome synthesis. Biochim Biophys Acta. 1985 Dec 18;826(4):167–173. doi: 10.1016/0167-4781(85)90002-8. [DOI] [PubMed] [Google Scholar]
  27. Yung B. Y., Chan P. K. Identification and characterization of a hexameric form of nucleolar phosphoprotein B23. Biochim Biophys Acta. 1987 Jul 16;925(1):74–82. doi: 10.1016/0304-4165(87)90149-8. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES