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
. 1990 May;87(9):3269–3273. doi: 10.1073/pnas.87.9.3269

Cellular levels of thymosin immunoreactive peptides are linked to proliferative events: evidence for a nuclear site of action.

C N Conteas 1, M G Mutchnick 1, K C Palmer 1, F E Weller 1, G D Luk 1, P H Naylor 1, M R Erdos 1, A L Goldstein 1, C Panneerselvam 1, B L Horecker 1
PMCID: PMC53881  PMID: 2375791

Abstract

Thymosin alpha 1 (T alpha 1), the N-terminal 28-amino acid fragment of prothymosin alpha (ProT alpha), and ProT alpha, although originally isolated from whole thymus extracts, are also present in nonthymic cells and tissues. We used an ELISA with an antibody raised against T alpha 1 to investigate the relationship between intracellular levels of thymosin immunoreactive peptide(s) (TIP) and cell proliferation in a rat small intestinal IEC-6 cell line. Increasing TIP levels were observed during cell proliferation, which decreased when proliferation was halted by cellular contact inhibition. Serum feeding of cells previously rendered quiescent by serum starvation resulted in a significant increase in TIP within 1 hr. Conversely, serum starvation decreased TIP levels within 1 hr. Peak TIP levels appeared after 3 hr of serum incubation, while maximum [3H]thymidine incorporation was noted after 9 hr, suggesting maximum TIP concentrations in the G1 phase of the proliferative cycle. Immunoelectron microscopy demonstrated an association of TIP with condensed nuclear chromatin. These results support a relation of intracellular TIP levels to IEC-6 cell proliferation and also a nuclear site of action. HPLC analysis of cellular homogenates from proliferating IEC-6 cells revealed a peak of immune reactivity that elutes in the position of T alpha 1.

Full text

PDF
3269

Images in this article

Selected References

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

  1. Conteas C. N., Majumdar A. P. The effects of gastrin, epidermal growth factor, and somatostatin on DNA synthesis in a small intestinal crypt cell line (IEC-6). Proc Soc Exp Biol Med. 1987 Mar;184(3):307–311. doi: 10.3181/00379727-184-42484. [DOI] [PubMed] [Google Scholar]
  2. Eschenfeldt W. H., Berger S. L. The human prothymosin alpha gene is polymorphic and induced upon growth stimulation: evidence using a cloned cDNA. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9403–9407. doi: 10.1073/pnas.83.24.9403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eschenfeldt W. H., Manrow R. E., Krug M. S., Berger S. L. Isolation and partial sequencing of the human prothymosin alpha gene family. Evidence against export of the gene products. J Biol Chem. 1989 May 5;264(13):7546–7555. [PubMed] [Google Scholar]
  4. Franco F. J., Diaz C., Barcia M., Arias P., Gomez-Marquez J., Soriano F., Mendez E., Freire M. Synthesis and apparent secretion of prothymosin alpha by different subpopulations of calf and rat thymocytes. Immunology. 1989 Jun;67(2):263–268. [PMC free article] [PubMed] [Google Scholar]
  5. Goldstein A. L., Low T. L., McAdoo M., McClure J., Thurman G. B., Rossio J., Lai C. Y., Chang D., Wang S. S., Harvey C. Thymosin alpha1: isolation and sequence analysis of an immunologically active thymic polypeptide. Proc Natl Acad Sci U S A. 1977 Feb;74(2):725–729. doi: 10.1073/pnas.74.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goodall G. J., Dominguez F., Horecker B. L. Molecular cloning of cDNA for human prothymosin alpha. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8926–8928. doi: 10.1073/pnas.83.23.8926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gómez-Márquez J., Segade F., Dosil M., Pichel J. G., Bustelo X. R., Freire M. The expression of prothymosin alpha gene in T lymphocytes and leukemic lymphoid cells is tied to lymphocyte proliferation. J Biol Chem. 1989 May 25;264(15):8451–8454. [PubMed] [Google Scholar]
  8. Gómez-Márquez J., Segade F. Prothymosin alpha is a nuclear protein. FEBS Lett. 1988 Jan 4;226(2):217–219. doi: 10.1016/0014-5793(88)81425-x. [DOI] [PubMed] [Google Scholar]
  9. Haritos A. A., Goodall G. J., Horecker B. L. Prothymosin alpha: isolation and properties of the major immunoreactive form of thymosin alpha 1 in rat thymus. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1008–1011. doi: 10.1073/pnas.81.4.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haritos A. A., Tsolas O., Horecker B. L. Distribution of prothymosin alpha in rat tissues. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1391–1393. doi: 10.1073/pnas.81.5.1391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hirokawa K., McClure J. E., Goldstein A. L. Age-related changes in localization of thymosin in the human thymus. Thymus. 1982 Jan;4(1):19–29. [PubMed] [Google Scholar]
  12. Kurokowa M., Lynch K., Podolsky D. K. Effects of growth factors on an intestinal epithelial cell line: transforming growth factor beta inhibits proliferation and stimulates differentiation. Biochem Biophys Res Commun. 1987 Feb 13;142(3):775–782. doi: 10.1016/0006-291x(87)91481-1. [DOI] [PubMed] [Google Scholar]
  13. Makarova T., Grebenshikov N., Egorov C., Vartapetian A., Bogdanov A. Prothymosin alpha is an evolutionary conserved protein covalently linked to a small RNA. FEBS Lett. 1989 Nov 6;257(2):247–250. doi: 10.1016/0014-5793(89)81544-3. [DOI] [PubMed] [Google Scholar]
  14. McClure J. E., Lameris N., Wara D. W., Goldstein A. L. Immunochemical studies on thymosin: radioimmunoassay of thymosin alpha 1. J Immunol. 1982 Jan;128(1):368–375. [PubMed] [Google Scholar]
  15. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  16. Panneerselvam C., Haritos A. A., Caldarella J., Horecker B. L. Prothymosin alpha in human blood. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4465–4469. doi: 10.1073/pnas.84.13.4465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Piltch A., Naylor P., Hayashi J. A cloned rat thymic epithelial cell line established from serum-free selective culture. In Vitro Cell Dev Biol. 1988 Apr;24(4):289–293. doi: 10.1007/BF02628829. [DOI] [PubMed] [Google Scholar]
  18. Quaroni A., May R. J. Establishment and characterizaton of intestinal epithelial cell cultures. Methods Cell Biol. 1980;21B:403–427. [PubMed] [Google Scholar]
  19. Quaroni A., Wands J., Trelstad R. L., Isselbacher K. J. Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria. J Cell Biol. 1979 Feb;80(2):248–265. doi: 10.1083/jcb.80.2.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wada S., Naylor P. H., Naylor C. W., Goldstein A. L. Improved ELISA to measure thymosin alpha 1: comparison of whole and absorbed antisera. Int J Immunopharmacol. 1988;10(7):795–801. doi: 10.1016/0192-0561(88)90002-1. [DOI] [PubMed] [Google Scholar]
  21. Watts J. D., Cary P. D., Crane-Robinson C. Prothymosin alpha is a nuclear protein. FEBS Lett. 1989 Mar 13;245(1-2):17–20. doi: 10.1016/0014-5793(89)80182-6. [DOI] [PubMed] [Google Scholar]
  22. Weller F. E., Mutchnick M. G., Keren D. F., Goldstein A. L., Naylor P. H. MicroELISA method for measurement of human serum thymosin alpha 1. J Immunol Methods. 1985 Jun 12;80(1):45–53. doi: 10.1016/0022-1759(85)90163-2. [DOI] [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