Letter | Published:

Na+/H+ exchanger regulatory factor 2 directs parathyroid hormone 1 receptor signalling

Naturevolume 417pages858861 (2002) | Download Citation

Subjects

Abstract

The parathyroid hormone 1 receptor (PTH1R) is a class II G-protein-coupled receptor1. PTH1R agonists include both PTH, a hormone that regulates blood calcium and phosphate, and PTH-related protein (PTHrP), a paracrine/autocrine factor that is essential for development, particularly of the skeleton. Adenylyl cyclase activation is thought to be responsible for most cellular responses to PTH and PTHrP, although many actions appear to be independent of adenylyl cyclase1,2,3,4,5. Here we show that the PTH1R binds to Na+/H+ exchanger regulatory factors (NHERF) 1 and 2 through a PDZ-domain interaction in vitro and in PTH target cells. NHERF2 simultaneously binds phospholipase Cβ1 and an atypical, carboxyl-terminal PDZ consensus motif, ETVM, of the PTH1R through PDZ1 and PDZ2, respectively. PTH treatment of cells that express the NHERF2–PTH1R complex markedly activates phospholipase Cβ and inhibits adenylyl cyclase through stimulation of inhibitory G proteins (Gi/o proteins). NHERF-mediated assembly of PTH1R and phospholipase Cβ is a unique mechanism to regulate PTH signalling in cells and membranes of polarized cells that express NHERF, which may account for many tissue- and cell-specific actions of PTH/PTHrP and may also be relevant to signalling by many G-protein-coupled receptors.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Nissenson, R. in The Parathyroids: Basic and Clinical Concepts, 2nd edn (eds Bilezikian, J., Marcus, R. & Levine, M.) 93–104 (Academic, San Diego, 2001)

  2. 2

    Strewler, G. J. in The Parathyroids: Basic and Clinical Concepts, 2nd edn (eds Bilezikian, J., Marcus, R. & Levine, M.) 213–226 (Academic, San Diego, 2001)

  3. 3

    Bringhurst, F. R. in The Parathyroids: Basic and Clinical Concepts, 2nd edn (eds Bilezikian, J., Marcus, R. & Levine, M.) 227–244 (Academic, San Diego, 2001)

  4. 4

    Segre, G. & Lee, K. in The Parathyroids: Basic and Clinical Concepts, 2nd edn (eds Bilezikian, J., Marcus, R. & Levine, M.) 245–260 (Academic, San Diego, 2001)

  5. 5

    Clemons, T. L. & Broadus, A. E. in The Parathyroids: Basic and Clinical Concepts, 2nd edn (eds Bilezikian, J., Marcus, R. & Levine, M.) 261–274 (Academic, San Diego, 2001)

  6. 6

    Abou-Samra, A. B. et al. Expression cloning of a common receptor for parathyroid hormone and parathyroid hormone-related peptide from rat osteoblast-like cells: A single receptor stimulates intracellular accumulation of both cAMP and inositol triphosphates and increases intracellular free calcium. Proc. Natl Acad. Sci. USA 89, 2732–2736 (1992)

  7. 7

    Reshkin, S. J., Forgo, J. & Murer, H. Apical and basolateral effects of PTH in OK cells: transport inhibition, messenger production, effects of pertussis toxin, and interaction with a PTH analog. J. Membr. Biol. 124, 227–237 (1991)

  8. 8

    Tanaka, H., Smogorzewski, M., Koss, M. & Massry, S. G. Pathways involved in PTH-induced rise in cytosolic Ca2+ concentration of rat renal proximal tubule. Am. J. Physiol. 268, F330–F337 (1995)

  9. 9

    Smogorzewski, M., Zayed, M., Zhang, Y. B., Roe, J. & Massry, S. G. Parathyroid hormone increases cytosolic calcium concentration in adult rat cardiac myocytes. Am. J. Physiol. 264, H1998–H2006 (1993)

  10. 10

    Miyauchi, A. et al. Stimulation of transient elevations in cytosolic Ca2+ is related to inhibition of Pi transport in OK cells. Am. J. Physiol. 259, F485–F493 (1990)

  11. 11

    Iida-Klein, A. et al. Truncation of the carboxyl-terminal region of the rat parathyroid hormone (PTH)/PTH-related peptide receptor enhances PTH stimulation of adenylyl cyclase but not phospholipase C. J. Biol. Chem. 270, 8458–8465 (1995)

  12. 12

    Doyle, D. A. et al. Crystal structures of a complexed and peptide-free membrane protein-binding domain: molecular basis of peptide recognition by PDZ. Cell 85, 1067–1076 (1996)

  13. 13

    Bretscher, A., Chambers, D., Nguyen, R. & Reczek, D. ERM-Merlin and EBP50 protein families in plasma membrane organization and function. Annu. Rev. Cell Dev. Biol. 16, 113–143 (2000)

  14. 14

    Yun, C. H. et al. cAMP-mediated inhibition of the epithelial brush border Na+/H+ exchanger, NHE3, requires an associated regulatory protein. Proc. Natl Acad. Sci. USA 94, 3010–3015 (1997)

  15. 15

    Krayer-Pawlowska, D., Helmle-Kolb, C., Montrose, M. H., Krapf, R. & Murer, H. Studies on the kinetics of Na+/H+ exchange in OK cells: introduction of a new device for the analysis of polarized transport in cultured epithelia. J. Membr. Biol. 120, 173–183 (1991)

  16. 16

    Hall, R. A. et al. A C-terminal motif found in the beta2-adrenergic receptor, P2Y1 receptor and cystic fibrosis transmembrane conductance regulator determines binding to the Na+/H+ exchanger regulatory factor family of PDZ proteins. Proc. Natl Acad. Sci. USA 95, 8496–8501 (1998)

  17. 17

    Wang, S., Raab, R. W., Schatz, P. J., Guggino, W. B. & Li, M. Peptide binding consensus of the NHE-RF-PDZ1 domain matches the C-terminal sequence of cystic fibrosis transmembrane conductance regulator (CFTR). FEBS Lett. 427, 103–108 (1998)

  18. 18

    Tang, Y. et al. Association of mammalian trp4 and phospholipase C isozymes with a PDZ domain-containing protein, NHERF. J. Biol. Chem. 275, 37559–37564 (2000)

  19. 19

    Amizuka, N. et al. Cell-specific expression of the parathyroid hormone (PTH)/PTH-related peptide receptor gene in kidney from kidney-specific and ubiquitous promoters. Endocrinology 138, 469–481 (1997)

  20. 20

    Wade, J. B., Welling, P. A., Donowitz, M., Shenolikar, S. & Weinman, E. J. Differential renal distribution of NHERF isoforms and their colocalization with NHE3, ezrin, and ROMK. Am. J. Physiol. Cell Physiol. 280, C192–C198 (2001)

  21. 21

    Isales, C. M. et al. Functional parathyroid hormone receptors are present in an umbilical vein endothelial cell line. Am. J. Physiol. Endocrinol. Metabol. 279, E654–E662 (2000)

  22. 22

    Yamamoto, I., Shigeno, C., Potts, J. T. Jr & Segre, G. V. Characterization and agonist-induced down-regulation of parathyroid hormone receptors in clonal rat osteosarcoma cells. Endocrinology 122, 1208–1217 (1988)

  23. 23

    Kaufmann, M. et al. Apical and basolateral parathyroid hormone receptors in rat renal cortical membranes. Endocrinology 134, 1173–1178 (1994)

  24. 24

    Traebert, M., Volkl, H., Biber, J., Murer, H. & Kaissling, B. Luminal and contraluminal action of 1-34 and 3-34 PTH peptides on renal type IIa Na-P(i) cotransporter. Am. J. Physiol. Renal Physiol. 278, F792–F798 (2000)

  25. 25

    Sutliff, R. L. et al. Vasorelaxant properties of parathyroid hormone-related protein in the mouse: evidence for endothelium involvement independent of nitric oxide formation. Endocrinology 140, 2077–2083 (1999)

  26. 26

    Orloff, J. J. et al. Analysis of PTHRP binding and signal transduction mechanisms in benign and malignant squamous cells. Am. J. Physiol. 262, E599–E607 (1992)

  27. 27

    Gaich, G. et al. Amino-terminal parathyroid hormone-related protein: specific binding and cytosolic calcium responses in rat insulinoma cells. Endocrinology 132, 1402–1409 (1993)

  28. 28

    Schluter, K. D., Weber, M. & Piper, H. M. Parathyroid hormone induces protein kinase C but not adenylate cyclase in adult cardiomyocytes and regulates cyclic AMP levels via protein kinase C-dependent phosphodiesterase activity. Biochem. J. 310, 439–444 (1995)

  29. 29

    McCauley, L. K., Rosol, T. J., Merryman, J. I. & Capen, C. C. Parathyroid hormone-related protein binding to human T-cell lymphotropic virus type I-infected lymphocytes. Endocrinology 130, 300–306 (1992)

  30. 30

    Tsunoda, S. et al. A multivalent PDZ-domain protein assembles signalling complexes in a G-protein-coupled cascade. Nature 388, 243–249 (1997)

  31. 31

    Kerjaschki, D. & Farquhar, M. G. Immunocytochemical localization of the Heymann nephritis antigen (GP330) in glomerular epithelial cells of normal Lewis rats. J. Exp. Med. 157, 667–686 (1983)

Download references

Acknowledgements

We thank D. Moore for providing the pT7-hismyc vector; S. Goo Rhee for providing the cDNA for PLCβ1; V. Ramesh for the NHERF1 cDNA; and C. Isales for providing the ECV304 cells. We also would like to thank J. Potts and H. Kronenberg for their helpful discussions and review of the manuscript. This work was supported in part by the NIH (G.V.S.).

Author information

Affiliations

  1. Endocrine Unit, Massachusetts General Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts, 02114, USA

    • Matthew J. Mahon
    •  & Gino V. Segre
  2. Department of Medicine, Gastroenterology Division, The Johns Hopkins University School of Medicine, Boston, Massachusetts, 21205, USA

    • Mark Donowitz
    •  & C. Chris Yun

Authors

  1. Search for Matthew J. Mahon in:

  2. Search for Mark Donowitz in:

  3. Search for C. Chris Yun in:

  4. Search for Gino V. Segre in:

Competing interests

The authors declare that they have no competing financial interests.

Corresponding author

Correspondence to Gino V. Segre.

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/nature00816

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.