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. 1977 Jun 1;73(3):761–767. doi: 10.1083/jcb.73.3.761

Increased ouabain-sensitive 86Rubidium uptake after mitogenic stimulation of quiescent chicken embryo fibroblasts with purified multiplication-stimulating activity

PMCID: PMC2111430  PMID: 141453

Abstract

Multiplication-stimulating activity (MSA), a protein which stimulates DNA synthesis and growth of chicken embryo fibroblasts, was purified from serum-free medium conditioned by the growth of a rat liver cell line. Purified MSA was shown to rapidly stimulate ouabain-sensitive Na+, K+-ATPase activity as measured by both enzyme assay and rate of 86Rubidium uptake. Labeled ouabain binding was also shown to increase after stimulation of quiescent cells by serum or purified MSA. Conditions which interfere with the ability of the cells to accumulate potassium, such as the presence of the specific inhibitor, ouabain; incubation in potassium-free medium; or the presence of the potassium ionophore, valinomycin, were all demonstrated to inhibit the stimulation of DNA synthesis by serum or purified MSA. These results suggest that an early event in the stimulation of DNA synthesis by purified MSA is an activation of membrane Na+, K+-ATPase with a resulting accumulation of potassium ions inside the cell.

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Selected References

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  1. Abaza N. A., Leighton J., Schultz S. G. Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney. I. Light microscopic observations of monolayer growth. In Vitro. 1974 Sep-Oct;10(3-4):72–183. doi: 10.1007/BF02615230. [DOI] [PubMed] [Google Scholar]
  2. Averdunk R., Lauf P. K. Effects of mitogens on sodium-potassium transport, 3H-ouabain binding, and adenosine triphosphatase activity in lymphocytes. Exp Cell Res. 1975 Jul;93(2):331–342. doi: 10.1016/0014-4827(75)90458-9. [DOI] [PubMed] [Google Scholar]
  3. Brodal B. P., Jebens E., Oy V., Iversen O. J. Effect of insulin on (Na+, K+)-activated adenosine triphosphatase activity in rat muscle sarcolemma. Nature. 1974 May 3;249(452):41–43. doi: 10.1038/249041a0. [DOI] [PubMed] [Google Scholar]
  4. Cone C. D., Jr, Tongier M., Jr Contact inhibition of division: involvement of the electrical transmembrane potential. J Cell Physiol. 1973 Dec;82(3):373–386. doi: 10.1002/jcp.1040820307. [DOI] [PubMed] [Google Scholar]
  5. Cuff J. M., Lichtman A. The early effects of ouabain on potassium metabolism and rate of proliferation of mouse lymphoblasts. J Cell Physiol. 1975 Apr;85(2 Pt 1):209–215. doi: 10.1002/jcp.1040850207. [DOI] [PubMed] [Google Scholar]
  6. Cuff J. M., Lichtman M. A. The effects of ouabain on the cell mitotic cycle of mouse lymphoblasts. J Cell Physiol. 1975 Apr;85(2 Pt 1):227–234. doi: 10.1002/jcp.1040850209. [DOI] [PubMed] [Google Scholar]
  7. Cunningham D. D., Pardee A. B. Transport changes rapidly initiated by serum addition to "contact inhibited" 3T3 cells. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1049–1056. doi: 10.1073/pnas.64.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dulak N. C., Temin H. M. A partially purified polypeptide fraction from rat liver cell conditioned medium with multiplication-stimulating activity for embryo fibroblasts. J Cell Physiol. 1973 Apr;81(2):153–160. doi: 10.1002/jcp.1040810204. [DOI] [PubMed] [Google Scholar]
  9. Dulak N. C., Temin H. M. Multiplication-stimulating activity for chicken embryo fibroblasts from rat liver cell conditioned medium: a family of small polypeptides. J Cell Physiol. 1973 Apr;81(2):161–170. doi: 10.1002/jcp.1040810205. [DOI] [PubMed] [Google Scholar]
  10. Holley R. W. A unifying hypothesis concerning the nature of malignant growth. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2840–2841. doi: 10.1073/pnas.69.10.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horn R. S., Walaas O., Walaas E. The influence of sodium, potassium and lithium on the response of glycogen synthetase I to insulin and epinephrine in the isolated rat diaphragm. Biochim Biophys Acta. 1973 Jul 28;313(2):296–309. doi: 10.1016/0304-4165(73)90029-9. [DOI] [PubMed] [Google Scholar]
  12. Kimelberg H. K., Mayhew E. Increased ouabain-sensitive 86Rb+ uptake and sodium and potassium ion-activated adenosine triphosphatase activity in transformed cell lines. J Biol Chem. 1975 Jan 10;250(1):100–104. [PubMed] [Google Scholar]
  13. Kimelberg H. K., Papahadjopoulos D. Phospholipid requirements for (Na + + K + )-ATPase activity: head-group specificity and fatty acid fluidity. Biochim Biophys Acta. 1972 Sep 1;282(1):277–292. doi: 10.1016/0005-2736(72)90334-3. [DOI] [PubMed] [Google Scholar]
  14. Kletzien R. F., Pariza M. W., Becker J. E., Potter V. R., Butcher F. R. Induction of amino acid transport in primary cultures of adult rat liver parenchymal cells by insulin. J Biol Chem. 1976 May 25;251(10):3014–3020. [PubMed] [Google Scholar]
  15. Kuchler R. J. The role of sodium and potassium in regulating amino acid accumulation and protein synthesis in LM-strain mouse fibroblasts. Biochim Biophys Acta. 1967 Apr 25;136(3):473–483. doi: 10.1016/0304-4165(67)90006-2. [DOI] [PubMed] [Google Scholar]
  16. Lostroh A. J., Krahl M. E. Insulin action. Accumulation in vitro of Mg 2 + and K + in rat uterus: ion pump activity. Biochim Biophys Acta. 1973 Jan 2;291(1):260–268. doi: 10.1016/0005-2736(73)90417-3. [DOI] [PubMed] [Google Scholar]
  17. Luly P., Barnabei O., Tria E. Hormonal control in vitro of plasma membrane-bound (Na + -K + )-ATPase of rat liver. Biochim Biophys Acta. 1972 Sep 1;282(1):447–452. doi: 10.1016/0005-2736(72)90352-5. [DOI] [PubMed] [Google Scholar]
  18. McDonald T. F., Sachs H. G., Orr C. W., Ebert J. D. Multiple effects of ouabain on BHK cells. Exp Cell Res. 1972 Sep;74(1):201–206. doi: 10.1016/0014-4827(72)90498-3. [DOI] [PubMed] [Google Scholar]
  19. Orr C. W., Yoshikawa-Fukada M., Ebert J. D. Potassium: effect on DNA synthesis and multiplication of baby-hamster kidney cells: (cell cycle-membrane potential-synchronization-transformation). Proc Natl Acad Sci U S A. 1972 Jan;69(1):243–247. doi: 10.1073/pnas.69.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Quastel M. R., Kaplan J. G. Early stimulation of potassium uptake in lymphocytes treated with PHA. Exp Cell Res. 1970 Nov;63(1):230–233. doi: 10.1016/0014-4827(70)90360-5. [DOI] [PubMed] [Google Scholar]
  21. Quastel M. R., Kaplan J. G. Lymphocyte stimulation: the effect of ouabain on nucleic acid and protein synthesis. Exp Cell Res. 1970 Oct;62(2):407–420. doi: 10.1016/0014-4827(70)90572-0. [DOI] [PubMed] [Google Scholar]
  22. Rozengurt E., Heppel L. A. Serum rapidly stimulates ouabain-sensitive 86-RB+ influx in quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4492–4495. doi: 10.1073/pnas.72.11.4492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rubin H., Koide T. Stimulation of DNA synthesis and 2-deoxy-D-glucose transport in chick embryo cultures by excessive metal concentrations and by a carcinogenic hydrocarbon. J Cell Physiol. 1973 Jun;81(3):387–396. doi: 10.1002/jcp.1040810311. [DOI] [PubMed] [Google Scholar]
  24. SCHATZMANN H. J. Herzglykoside als Hemmstoffe für den aktiven Kalium- und Natriumtransport durch die Erythrocytenmembran. Helv Physiol Pharmacol Acta. 1953;11(4):346–354. [PubMed] [Google Scholar]
  25. SKOU J. C. ENZYMATIC BASIS FOR ACTIVE TRANSPORT OF NA+ AND K+ ACROSS CELL MEMBRANE. Physiol Rev. 1965 Jul;45:596–617. doi: 10.1152/physrev.1965.45.3.596. [DOI] [PubMed] [Google Scholar]
  26. Schultz S. G., Curran P. F. Coupled transport of sodium and organic solutes. Physiol Rev. 1970 Oct;50(4):637–718. doi: 10.1152/physrev.1970.50.4.637. [DOI] [PubMed] [Google Scholar]
  27. Shank B. B., Smith N. E. Regulation of cellular growth by sodium pump activity. J Cell Physiol. 1976 Mar;87(3):377–387. doi: 10.1002/jcp.1040870313. [DOI] [PubMed] [Google Scholar]
  28. Shaw S. N., Amos H. Insulin stimulation of glucose entry in chick fibroblasts and HeLa cells. Biochem Biophys Res Commun. 1973 Jul 2;53(1):357–365. doi: 10.1016/0006-291x(73)91441-1. [DOI] [PubMed] [Google Scholar]
  29. Smith G. L., Temin H. M. Purified multiplication-stimulating activity from rat liver cell conditioned medium: comparison of biological activities with calf serum, insulin, and somatomedin. J Cell Physiol. 1974 Oct;84(2):181–192. doi: 10.1002/jcp.1040840204. [DOI] [PubMed] [Google Scholar]
  30. Vaughan G. L., Cook J. S. Regeneration of cation-transport capacity in HeLa cell membranes after specific blockade by ouabain. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2627–2631. doi: 10.1073/pnas.69.9.2627. [DOI] [PMC free article] [PubMed] [Google Scholar]

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