Skip to main content
PMC home page
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
. 1982 Oct;79(20):6275–6279. doi: 10.1073/pnas.79.20.6275

Intracellular K+ and the mitogenic response of 3T3 cells to peptide factors in serum-free medium.

A Lopez-Rivas, E A Adelberg, E Rozengurt
PMCID: PMC347103  PMID: 6755467

Abstract

The stimulation of DNA synthesis in cultures of Swiss 3T3 cells by vasopressin, epidermal growth factor, and insulin added in serum-free medium is strikingly dependent on the intracellular K+ content or concentration. The relationship between these parameters is sigmoid; DNA synthesis commences only when the intracellular K+ increases above a certain threshold level (0.56 mumol/mg of protein; 90 mM). Addition of K+ to K+- depleted cultures reverses the block on DNA synthesis after a lag period of at least 8 hr. The sigmoid dependence of DNA synthesis on intracellular K+ is generated in early G1 phase rather than at the G1/S boundary. The effects of K+ on the G1-S transition are, at least in part, exerted through its control of protein synthesis. In serum-free medium, the K+ content is close to the threshold required for allowing a mitogenic response. The findings suggest that a small change in the intracellular K+ level can influence the ability of these cells to initiate DNA synthesis in serum-free medium.

Full text

PDF
6276

Selected References

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

  1. Bourne H. R., Rozengurt E. An 18,000 molecular weight polypeptide induces early events and stimulates DNA synthesis in cultured cells. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4555–4559. doi: 10.1073/pnas.73.12.4555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brendler T., Godefroy-Colburn T., Yu S., Thach R. E. The role of mRNA competition in regulating translation. III. Comparison of in vitro and in vivo results. J Biol Chem. 1981 Nov 25;256(22):11755–11761. [PubMed] [Google Scholar]
  3. Brooks R. F. Continuous protein synthesis is required to maintain the probability of entry into S phase. Cell. 1977 Sep;12(1):311–317. doi: 10.1016/0092-8674(77)90209-4. [DOI] [PubMed] [Google Scholar]
  4. Cahn F., Lubin M. Inhibition of elongation steps of protein synthesis at reduced potassium concentrations in reticulocytes and reticulocyte lysate. J Biol Chem. 1978 Nov 10;253(21):7798–7803. [PubMed] [Google Scholar]
  5. Dicker P., Rozengurt E. Phorbol ester stimulation of Na influx and Na-K pump activity in Swiss 3T3 cells. Biochem Biophys Res Commun. 1981 May 15;100(1):433–441. doi: 10.1016/s0006-291x(81)80115-5. [DOI] [PubMed] [Google Scholar]
  6. Frantz C. N., Nathan D. G., Scher C. D. Intracellular univalent cations and the regulation of the BALB/c-3T3 cell cycle. J Cell Biol. 1981 Jan;88(1):51–56. doi: 10.1083/jcb.88.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kaplan P. L., Topp W. C., Ozanne B. Simian virus 40 induces the production of a polypeptide transforming factor(s). Virology. 1981 Jan 30;108(2):484–490. doi: 10.1016/0042-6822(81)90455-4. [DOI] [PubMed] [Google Scholar]
  8. Kletzien R. F., Pariza M. W., Becker J. E., Potter V. R. A method using 3-O-methyl-D-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture. Anal Biochem. 1975 Oct;68(2):537–544. doi: 10.1016/0003-2697(75)90649-1. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Ledbetter M. L., Lubin M. Control of protein synthesis in human fibroblasts by intracellular potassium. Exp Cell Res. 1977 Mar 15;105(2):223–236. doi: 10.1016/0014-4827(77)90120-3. [DOI] [PubMed] [Google Scholar]
  11. Lubin M. Control of growth by intracellular potassium and sodium concentrations is relaxed in transformed 3T3 cells. Biochem Biophys Res Commun. 1980 Dec 16;97(3):1060–1067. doi: 10.1016/0006-291x(80)91483-7. [DOI] [PubMed] [Google Scholar]
  12. Mendoza S. A., Wigglesworth N. M., Pohjanpelto P., Rozengurt E. Na entry and Na-K pump activity in murine, hamster, and human cells--effect of monensin, serum, platelet extract, and viral transformation. J Cell Physiol. 1980 Apr;103(1):17–27. doi: 10.1002/jcp.1041030104. [DOI] [PubMed] [Google Scholar]
  13. Mendoza S. A., Wigglesworth N. M., Rozengurt E. Vasopressin rapidly stimulates Na entry and Na-K pump activity in quiescent cultures of mouse 3T3 cells. J Cell Physiol. 1980 Oct;105(1):153–162. doi: 10.1002/jcp.1041050117. [DOI] [PubMed] [Google Scholar]
  14. Mierzejewski K., Rozengurt E. Vitamin B12 enhances the stimulation of DNA synthesis by serum in resting cultures of 3T6 cells. Exp Cell Res. 1977 May;106(2):394–397. doi: 10.1016/0014-4827(77)90187-2. [DOI] [PubMed] [Google Scholar]
  15. Paris S., Rozengurt E. Cyclic AMP stimulation of Na-K pump activity in quiescent swiss 3T3 cells. J Cell Physiol. 1982 Aug;112(2):273–280. doi: 10.1002/jcp.1041120217. [DOI] [PubMed] [Google Scholar]
  16. Pollack L. R., Tate E. H., Cook J. S. Turnover and regulation of Na-K-ATPase in HeLa cells. Am J Physiol. 1981 Nov;241(5):C173–C183. doi: 10.1152/ajpcell.1981.241.5.C173. [DOI] [PubMed] [Google Scholar]
  17. Rossow P. W., Riddle V. G., Pardee A. B. Synthesis of labile, serum-dependent protein in early G1 controls animal cell growth. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4446–4450. doi: 10.1073/pnas.76.9.4446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rozengurt E., Gelehrter T. D., Legg A., Pettican P. Melittin stimulates Na entry, Na-K pump activity and DNA synthesis in quiescent cultures of mouse cells. Cell. 1981 Mar;23(3):781–788. doi: 10.1016/0092-8674(81)90442-6. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Rozengurt E., Legg A., Pettican P. Vasopressin stimulation of mouse 3T3 cell growth. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1284–1287. doi: 10.1073/pnas.76.3.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rozengurt E., Legg A., Strang G., Courtenay-Luck N. Cyclic AMP: a mitogenic signal for Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4392–4396. doi: 10.1073/pnas.78.7.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rozengurt E. Stimulation of DNA synthesis in quiescent cultured cells: exogenous agents, internal signals, and early events. Curr Top Cell Regul. 1980;17:59–88. doi: 10.1016/b978-0-12-152817-1.50007-9. [DOI] [PubMed] [Google Scholar]
  23. Schneiderman M. H., Dewey W. C., Highfield D. P. Inhibition of DNA synthesis in synchronized Chinese hamster cells treated in G1 with cycloheximide. Exp Cell Res. 1971 Jul;67(1):147–155. doi: 10.1016/0014-4827(71)90630-6. [DOI] [PubMed] [Google Scholar]
  24. Smith J. B., Rozengurt E. Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5560–5564. doi: 10.1073/pnas.75.11.5560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tupper J. T., Zorgniotti F., Mills B. Potassium transport and content during G1 and S phase following serum stimulation of 3T3 cells. J Cell Physiol. 1977 Jun;91(3):429–440. doi: 10.1002/jcp.1040910313. [DOI] [PubMed] [Google Scholar]
  27. Wallick E. T., Lane L. K., Schwartz A. Biochemical mechanism of the sodium pump. Annu Rev Physiol. 1979;41:397–411. doi: 10.1146/annurev.ph.41.030179.002145. [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