Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1974 Oct 1;63(1):288–294. doi: 10.1083/jcb.63.1.288

CYCLIC AMP MEDIATES THE CONCANAVALIN A AGGLUTINABILITY OF MOUSE FIBROBLASTS

Mark C Willingham 1, Ira Pastan 1
PMCID: PMC2109349  PMID: 4371073

Abstract

We have devised a quantitative way to measure the agglutination of cells which utilizes the size discrimination feature of an automatic particle counter. With this method we have studied the agglutinability by concanavalin A of 3T3 cells, a mutant of 3T3 cells (3T3cAMPtcs) in which cyclic AMP levels fall when the cells are subjected to temperature change or fresh serum, and L929 cells. We find with 3T3cAMPtcs cells that low levels of cyclic AMP correlate with increased agglutinability and that high levels of cyclic AMP correlate with decreased agglutinability. Prior treatment of these cells with a cyclic AMP phosphodiesterase inhibitor or Bt2cAMP blocks the increase in agglutinability induced by temperature change. When 3T3 cells are treated with fresh serum, their agglutinability also increases although to a much smaller extent than with 3T3cAMPtcs cells. Cells change their agglutinability very rapidly. Treatment of L929 cells for 15 min with 1-methyl-3-isobutyl xanthine at 1 mM decreases their agglutinability to the level of normal 3T3 cells. We conclude that in normal and transformed cells the level of cyclic AMP regulates agglutinability.

Full Text

The Full Text of this article is available as a PDF (523.3 KB).

Selected References

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

  1. Burger M. M. A difference in the architecture of the surface membrane of normal and virally transformed cells. Proc Natl Acad Sci U S A. 1969 Mar;62(3):994–1001. doi: 10.1073/pnas.62.3.994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burger M. M., Bombik B. M., Breckenridge B. M., Sheppard J. R. Growth control and cyclic alterations of cyclic AMP in the cell cycle. Nat New Biol. 1972 Oct 11;239(93):161–163. doi: 10.1038/newbio239161a0. [DOI] [PubMed] [Google Scholar]
  3. Burger M. M., Goldberg A. R. Identification of a tumor-specific determinant on neoplastic cell surfaces. Proc Natl Acad Sci U S A. 1967 Feb;57(2):359–366. doi: 10.1073/pnas.57.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eckhart W., Dulbecco R., Burger M. M. Temperature-dependent surface changes in cells infected or transformed by a thermosensitive mutant of polyoma virus. Proc Natl Acad Sci U S A. 1971 Feb;68(2):283–286. doi: 10.1073/pnas.68.2.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hsie A. W., Jones C., Puck T. T. Further changes in differentiation state accompanying the conversion of Chinese hamster cells of fibroblastic form by dibutyryl adenosine cyclic 3':5'-monophosphate and hormones. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1648–1652. doi: 10.1073/pnas.68.7.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hsie A. W., Puck T. T. Morphological transformation of Chinese hamster cells by dibutyryl adenosine cyclic 3':5'-monophosphate and testosterone. Proc Natl Acad Sci U S A. 1971 Feb;68(2):358–361. doi: 10.1073/pnas.68.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Inbar M., Ben-Bassat H., Sachs L. A specific metabolic activity on the surface membrane in malignant cell-transformation. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2748–2751. doi: 10.1073/pnas.68.11.2748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Inbar M., Ben-Bassat H., Sachs L. Membrane changes associated with malignancy. Nat New Biol. 1972 Mar 1;236(61):3–passim. doi: 10.1038/newbio236003a0. [DOI] [PubMed] [Google Scholar]
  9. Inbar M., Sachs L. Interaction of the carbohydrate-binding protein concanavalin A with normal and transformed cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1418–1425. doi: 10.1073/pnas.63.4.1418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson G. S., Friedman R. M., Pastan I. Cyclic AMP-treated sarcoma cells acquire several morphological characteristics of normal fibroblasts. Ann N Y Acad Sci. 1971 Dec 30;185:413–416. doi: 10.1111/j.1749-6632.1971.tb45267.x. [DOI] [PubMed] [Google Scholar]
  11. Johnson G. S., Friedman R. M., Pastan I. Restoration of several morphological characteristics of normal fibroblasts in sarcoma cells treated with adenosine-3':5'-cyclic monphosphate and its derivatives. Proc Natl Acad Sci U S A. 1971 Feb;68(2):425–429. doi: 10.1073/pnas.68.2.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson G. S., Pastan I. Role of 3',5'-adenosine monophosphate in regulation of morphology and growth of transformed and normal fibroblasts. J Natl Cancer Inst. 1972 May;48(5):1377–1387. [PubMed] [Google Scholar]
  13. Kurth R., Bauer H. Influence of dibutyryl cyclicAMP and theophylline on cell surface antigens on oncornavirus transformed cells. Nat New Biol. 1973 Jun 20;243(129):243–245. doi: 10.1038/newbio243243a0. [DOI] [PubMed] [Google Scholar]
  14. Nicolson G. L. Temperature-dependent mobility of concanavalin A sites on tumour cell surfaces. Nat New Biol. 1973 Jun 13;243(128):218–220. doi: 10.1038/newbio243218a0. [DOI] [PubMed] [Google Scholar]
  15. Nicolson G. L. Topography of membrane concanavalin A sites modified by proteolysis. Nat New Biol. 1972 Oct 18;239(94):193–197. doi: 10.1038/newbio239193a0. [DOI] [PubMed] [Google Scholar]
  16. O'Neill C. H. Growth induction by serum or polyoma virus inhibits the aggregation of trypsinised suspensions of BHK21 tissue culture fibroblasts. Exp Cell Res. 1973 Sep;81(1):31–39. doi: 10.1016/0014-4827(73)90107-9. [DOI] [PubMed] [Google Scholar]
  17. Otten J., Bader J., Johnson G. S., Pastan I. A mutation in a rous sarcoma virus gene that controls adenosine 3',5'-monophosphate levels and transformation. J Biol Chem. 1972 Mar 10;247(5):1632–1633. [PubMed] [Google Scholar]
  18. Otten J., Johnson G. S., Pastan I. Cyclic AMP levels in fibroblasts: relationship to growth rate and contact inhibition of growth. Biochem Biophys Res Commun. 1971 Sep;44(5):1192–1198. doi: 10.1016/s0006-291x(71)80212-7. [DOI] [PubMed] [Google Scholar]
  19. Otten J., Johnson G. S., Pastan I. Regulation of cell growth by cyclic adenosine 3',5'-monophosphate. Effect of cell density and agents which alter cell growth on cyclic adenosine 3',5'-monophosphate levels in fibroblasts. J Biol Chem. 1972 Nov 10;247(21):7082–7087. [PubMed] [Google Scholar]
  20. Porter K., Prescott D., Frye J. Changes in surface morphology of Chinese hamster ovary cells during the cell cycle. J Cell Biol. 1973 Jun;57(3):815–836. doi: 10.1083/jcb.57.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rein A., Carchman R. A., Johnson G. S., Pastan I. Simian virus 40 rapidly lowers cAMP levels in mouse cells. Biochem Biophys Res Commun. 1973 Jun 8;52(3):899–904. doi: 10.1016/0006-291x(73)91022-x. [DOI] [PubMed] [Google Scholar]
  22. Sharon N., Lis H. Lectins: cell-agglutinating and sugar-specific proteins. Science. 1972 Sep 15;177(4053):949–959. doi: 10.1126/science.177.4053.949. [DOI] [PubMed] [Google Scholar]
  23. Sheppard J. R. Difference in the cyclic adenosine 3',5'-monophosphate levels in normal and transformed cells. Nat New Biol. 1972 Mar 1;236(61):14–16. doi: 10.1038/newbio236014a0. [DOI] [PubMed] [Google Scholar]
  24. Sheppard J. R., Prescott D. M. Cyclic AMP levels in synchronized mammalian cells. Exp Cell Res. 1972 Nov;75(1):293–296. doi: 10.1016/0014-4827(72)90554-x. [DOI] [PubMed] [Google Scholar]
  25. Sheppard J. R. Restoration of contact-inhibited growth to transformed cells by dibutyryl adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1316–1320. doi: 10.1073/pnas.68.6.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smets L. A. Contact inhibition of transformed cells incompletely restored by dibutyryl cyclic AMP. Nat New Biol. 1972 Sep 27;239(91):123–124. doi: 10.1038/newbio239123a0. [DOI] [PubMed] [Google Scholar]
  27. Thion C., Green M. Cyclic AMP-amplified replication of RNA tumour virus-like particles in Chinese hamster ovary cells. Nat New Biol. 1973 Aug 22;244(138):227–231. doi: 10.1038/newbio244227a0. [DOI] [PubMed] [Google Scholar]
  28. Willingham M. C., Carchman R. A., Pastan I. H. A mutant of 3T3 cells with cyclic AMP metabolism sensitive to temperature change. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2906–2910. doi: 10.1073/pnas.70.10.2906. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES