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
. 1992 Mar 15;89(6):2424–2428. doi: 10.1073/pnas.89.6.2424

The Na+/H+ antiporter cytoplasmic domain mediates growth factor signals and controls "H(+)-sensing".

S Wakabayashi 1, P Fafournoux 1, C Sardet 1, J Pouysségur 1
PMCID: PMC48670  PMID: 1372444

Abstract

The amiloride-sensitive Na+/H+ exchanger (NHE1 human isoform) is activated in response to diverse mitogenic and oncogenic signals presumably through phosphorylation. To get insight into the activating mechanism, a set of deletion mutants within the C-terminal cytoplasmic domain of NHE1 has been generated. These mutant forms expressed in antiporter-deficient fibroblasts revealed that deletion of the complete cytoplasmic domain (i) preserves amiloride-sensitive Na+/H+ exchange and activation by intracellular H+, (ii) reduces the affinity of the internal "H(+)-modifier site" in a manner mimicked by cellular ATP depletion, and (iii) abolishes growth factor-induced cytoplasmic alkalinization. We conclude that NHE1 can be separated into two distinct functional domains. One is an N-terminal transporter domain (T) that has all the features required to catalyze amiloride-sensitive Na+/H+ exchange with a built-in H(+)-modifier site. The other is a C-terminal cytoplasmic regulatory domain (R) that (i) determines the set point value of the exchanger and (ii) mediates growth factor signals by interacting with the "H(+)-sensor" in a phosphorylation-dependent manner.

Full text

PDF
2424

Images in this article

2425Image
on p.2425
2426Image
on p.2426
2426Image
on p.2426
2427Image
on p.2427
2428Image
on p.2428

Selected References

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

  1. Aronson P. S., Nee J., Suhm M. A. Modifier role of internal H+ in activating the Na+-H+ exchanger in renal microvillus membrane vesicles. Nature. 1982 Sep 9;299(5879):161–163. doi: 10.1038/299161a0. [DOI] [PubMed] [Google Scholar]
  2. Cassel D., Katz M., Rotman M. Depletion of cellular ATP inhibits Na+/H+ antiport in cultured human cells. Modulation of the regulatory effect of intracellular protons on the antiporter activity. J Biol Chem. 1986 Apr 25;261(12):5460–5466. [PubMed] [Google Scholar]
  3. Ellis L., Clauser E., Morgan D. O., Edery M., Roth R. A., Rutter W. J. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose. Cell. 1986 Jun 6;45(5):721–732. doi: 10.1016/0092-8674(86)90786-5. [DOI] [PubMed] [Google Scholar]
  4. Franchi A., Cragoe E., Jr, Pouysségur J. Isolation and properties of fibroblast mutants overexpressing an altered Na+/H+ antiporter. J Biol Chem. 1986 Nov 5;261(31):14614–14620. [PubMed] [Google Scholar]
  5. Ganz M. B., Boyarsky G., Sterzel R. B., Boron W. F. Arginine vasopressin enhances pHi regulation in the presence of HCO3- by stimulating three acid-base transport systems. Nature. 1989 Feb 16;337(6208):648–651. doi: 10.1038/337648a0. [DOI] [PubMed] [Google Scholar]
  6. Grinstein S., Rotin D., Mason M. J. Na+/H+ exchange and growth factor-induced cytosolic pH changes. Role in cellular proliferation. Biochim Biophys Acta. 1989 Jan 18;988(1):73–97. doi: 10.1016/0304-4157(89)90004-x. [DOI] [PubMed] [Google Scholar]
  7. L'Allemain G., Paris S., Pouysségur J. Growth factor action and intracellular pH regulation in fibroblasts. Evidence for a major role of the Na+/H+ antiport. J Biol Chem. 1984 May 10;259(9):5809–5815. [PubMed] [Google Scholar]
  8. L'Allemain G., Paris S., Pouysségur J. Role of a Na+-dependent Cl-/HCO3- exchange in regulation of intracellular pH in fibroblasts. J Biol Chem. 1985 Apr 25;260(8):4877–4883. [PubMed] [Google Scholar]
  9. Maly K., Uberall F., Loferer H., Doppler W., Oberhuber H., Groner B., Grunicke H. H. Ha-ras activates the Na+/H+ antiporter by a protein kinase C-independent mechanism. J Biol Chem. 1989 Jul 15;264(20):11839–11842. [PubMed] [Google Scholar]
  10. Margolis L. B., Rozovskaja I. A., Cragoe E. Intracellular pH and cell adhesion to solid substrate. FEBS Lett. 1988 Jul 18;234(2):449–450. doi: 10.1016/0014-5793(88)80135-2. [DOI] [PubMed] [Google Scholar]
  11. Meloche S., Pagès G., Pouysségur J. Functional expression and growth factor activation of an epitope-tagged p44 mitogen-activated protein kinase, p44mapk. Mol Biol Cell. 1992 Jan;3(1):63–71. doi: 10.1091/mbc.3.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Moolenaar W. H. Effects of growth factors on intracellular pH regulation. Annu Rev Physiol. 1986;48:363–376. doi: 10.1146/annurev.ph.48.030186.002051. [DOI] [PubMed] [Google Scholar]
  13. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  14. Paris S., Pouysségur J. Growth factors activate the Na+/H+ antiporter in quiescent fibroblasts by increasing its affinity for intracellular H+. J Biol Chem. 1984 Sep 10;259(17):10989–10994. [PubMed] [Google Scholar]
  15. Pouysségur J., Sardet C., Franchi A., L'Allemain G., Paris S. A specific mutation abolishing Na+/H+ antiport activity in hamster fibroblasts precludes growth at neutral and acidic pH. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4833–4837. doi: 10.1073/pnas.81.15.4833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sardet C., Counillon L., Franchi A., Pouysségur J. Growth factors induce phosphorylation of the Na+/H+ antiporter, glycoprotein of 110 kD. Science. 1990 Feb 9;247(4943):723–726. doi: 10.1126/science.2154036. [DOI] [PubMed] [Google Scholar]
  17. Sardet C., Fafournoux P., Pouysségur J. Alpha-thrombin, epidermal growth factor, and okadaic acid activate the Na+/H+ exchanger, NHE-1, by phosphorylating a set of common sites. J Biol Chem. 1991 Oct 15;266(29):19166–19171. [PubMed] [Google Scholar]
  18. Sardet C., Franchi A., Pouysségur J. Molecular cloning, primary structure, and expression of the human growth factor-activatable Na+/H+ antiporter. Cell. 1989 Jan 27;56(2):271–280. doi: 10.1016/0092-8674(89)90901-x. [DOI] [PubMed] [Google Scholar]
  19. Schwartz M. A., Both G., Lechene C. Effect of cell spreading on cytoplasmic pH in normal and transformed fibroblasts. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4525–4529. doi: 10.1073/pnas.86.12.4525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tse C. M., Ma A. I., Yang V. W., Watson A. J., Levine S., Montrose M. H., Potter J., Sardet C., Pouyssegur J., Donowitz M. Molecular cloning and expression of a cDNA encoding the rabbit ileal villus cell basolateral membrane Na+/H+ exchanger. EMBO J. 1991 Aug;10(8):1957–1967. doi: 10.1002/j.1460-2075.1991.tb07725.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Van Obberghen-Schilling E., Chambard J. C., Paris S., L'Allemain G., Pouysségur J. alpha-Thrombin-induced early mitogenic signalling events and G0 to S-phase transition of fibroblasts require continual external stimulation. EMBO J. 1985 Nov;4(11):2927–2932. doi: 10.1002/j.1460-2075.1985.tb04025.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wilson I. A., Niman H. L., Houghten R. A., Cherenson A. R., Connolly M. L., Lerner R. A. The structure of an antigenic determinant in a protein. Cell. 1984 Jul;37(3):767–778. doi: 10.1016/0092-8674(84)90412-4. [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