Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Modification of mineralocorticoid receptor function by Rac1 GTPase: implication in proteinuric kidney disease

Nature Medicine volume 14, pages 1370–1376 (2008)Cite this article

Abstract

Blockade of mineralocorticoid receptor has been shown to improve the clinical outcomes of proteinuric kidney diseases1,2. However, little is known about the regulation of mineralocorticoid receptor–dependent transcriptional activity in renal disease. Here we identify a new role for Rac1, a member of the Rho family GTPases, as a potent activator of mineralocorticoid receptor signal transduction both in vitro and in vivo. Transient transfection assays in HEK 293 cells revealed that constitutively active Rac1 (CA-Rac1) enhanced mineralocorticoid receptor–dependent reporter activity, which was accompanied by increased nuclear translocation of mineralocorticoid receptor. CA-Rac1 facilitated mineralocorticoid receptor nuclear accumulation also in podocytes via p21-activated kinase phosphorylation. In mice lacking Rho GDP-dissociation inhibitor-α (Arhgdia−/− mice)3, renal abnormalities, including heavy albuminuria and podocyte damage, were associated with increased Rac1 (but not RhoA) and mineralocorticoid receptor signaling in the kidney, without alteration in systemic aldosterone status. Pharmacological intervention with a Rac-specific small-molecule inhibitor4,5 diminished mineralocorticoid receptor overactivity and renal damage in this model. Furthermore, albuminuria and histological changes in Arhgdia−/− mice were suppressed by mineralocorticoid receptor blockade, confirming the pathological role of Rac1-mineralocorticoid receptor interaction. Our results provide evidence that signaling cross-talk between Rac1 and mineralocorticoid receptor modulates mineralocorticoid receptor activity and identify Rac1 as a therapeutic target for chronic kidney disease.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Active Rac1 enhances mineralocorticoid receptor (MR) transcriptional activity and nuclear translocation.
Figure 2: Renal damage, activity of Rho family GTPases and MR signaling in Arhgdia−/− mice.
Figure 3: Effects of NSC23766 on renal damage of Arhgdia−/− mice.
Figure 4: MR signaling mediates albuminuria and renal injury in Arhgdia−/− mice.

Similar content being viewed by others

References

  1. Chrysostomou, A. & Becker, G. Spironolactone in addition to ACE inhibition to reduce proteinuria in patients with chronic renal disease. N. Engl. J. Med. 345, 925–926 (2001).

    Article  CAS  Google Scholar 

  2. Williams, G.H. et al. Efficacy of eplerenone versus enalapril as monotherapy in systemic hypertension. Am. J. Cardiol. 93, 990–996 (2004).

    Article  CAS  Google Scholar 

  3. Togawa, A. et al. Progressive impairment of kidneys and reproductive organs in mice lacking Rho GDIalpha. Oncogene 18, 5373–5380 (1999).

    Article  CAS  Google Scholar 

  4. Gao, Y., Dickerson, J.B., Guo, F., Zheng, J. & Zheng, Y. Rational design and characterization of a Rac GTPase–specific small molecule inhibitor. Proc. Natl Acad. Sci. USA 101, 7618–7623 (2004).

    Article  CAS  Google Scholar 

  5. Cancelas, J.A. et al. Rac GTPases differentially integrate signals regulating hematopoietic stem cell localization. Nat. Med. 11, 886–891 (2005).

    Article  CAS  Google Scholar 

  6. Kato, S. et al. Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase. Science 270, 1491–1494 (1995).

    Article  CAS  Google Scholar 

  7. Tronche, F. et al. Glucocorticoid receptor function in hepatocytes is essential to promote postnatal body growth. Genes Dev. 18, 492–497 (2004).

    Article  CAS  Google Scholar 

  8. Takai, Y., Sasaki, T. & Matozaki, T. Small GTP-binding proteins. Physiol. Rev. 81, 153–208 (2001).

    Article  CAS  Google Scholar 

  9. Tzima, E. et al. Activation of Rac1 by shear stress in endothelial cells mediates both cytoskeletal reorganization and effects on gene expression. EMBO J. 21, 6791–6800 (2002).

    Article  CAS  Google Scholar 

  10. Rubino, D. et al. Characterization of Brx, a novel Dbl family member that modulates estrogen receptor action. Oncogene 16, 2513–2526 (1998).

    Article  CAS  Google Scholar 

  11. Su, L.F., Knoblauch, R. & Garabedian, M.J. Rho GTPases as modulators of the estrogen receptor transcriptional response. J. Biol. Chem. 276, 3231–3237 (2001).

    Article  CAS  Google Scholar 

  12. Kino, T. et al. Rho family Guanine nucleotide exchange factor Brx couples extracellular signals to the glucocorticoid signaling system. J. Biol. Chem. 281, 9118–9126 (2006).

    Article  CAS  Google Scholar 

  13. Le Menuet, D. et al. Alteration of cardiac and renal functions in transgenic mice overexpressing human mineralocorticoid receptor. J. Biol. Chem. 276, 38911–38920 (2001).

    Article  CAS  Google Scholar 

  14. Massaad, C., Houard, N., Lombes, M. & Barouki, R. Modulation of human mineralocorticoid receptor function by protein kinase A. Mol. Endocrinol. 13, 57–65 (1999).

    Article  CAS  Google Scholar 

  15. Funder, J.W. Is aldosterone bad for the heart? Trends Endocrinol. Metab. 15, 139–142 (2004).

    Article  CAS  Google Scholar 

  16. Nagase, M., Matsui, H., Shibata, S., Gotoda, T. & Fujita, T. Salt-induced nephropathy in obese spontaneously hypertensive rats via paradoxical activation of the mineralocorticoid receptor: role of oxidative stress. Hypertension 50, 877–883 (2007).

    Article  CAS  Google Scholar 

  17. Burridge, K. & Wennerberg, K. Rho and Rac take center stage. Cell 116, 167–179 (2004).

    Article  CAS  Google Scholar 

  18. Edwards, D.C., Sanders, L.C., Bokoch, G.M. & Gill, G.N. Activation of LIM-kinase by Pak1 couples Rac/Cdc42 GTPase signalling to actin cytoskeletal dynamics. Nat. Cell Biol. 1, 253–259 (1999).

    Article  CAS  Google Scholar 

  19. Wang, R.A., Mazumdar, A., Vadlamudi, R.K. & Kumar, R. P21-activated kinase-1 phosphorylates and transactivates estrogen receptor-alpha and promotes hyperplasia in mammary epithelium. EMBO J. 21, 5437–5447 (2002).

    Article  CAS  Google Scholar 

  20. Zhao, L. et al. Role of p21-activated kinase pathway defects in the cognitive deficits of Alzheimer disease. Nat. Neurosci. 9, 234–242 (2006).

    Article  CAS  Google Scholar 

  21. Shibata, S., Nagase, M. & Fujita, T. Fluvastatin ameliorates podocyte injury in proteinuric rats via modulation of excessive Rho signaling. J. Am. Soc. Nephrol. 17, 754–764 (2006).

    Article  CAS  Google Scholar 

  22. Shibata, S., Nagase, M., Yoshida, S., Kawachi, H. & Fujita, T. Podocyte as the target for aldosterone: roles of oxidative stress and Sgk1. Hypertension 49, 355–364 (2007).

    Article  CAS  Google Scholar 

  23. Nagase, M. et al. Podocyte injury underlies the glomerulopathy of Dahl salt-hypertensive rats and is reversed by aldosterone blocker. Hypertension 47, 1084–1093 (2006).

    Article  CAS  Google Scholar 

  24. Brown, N.J. et al. Aldosterone modulates plasminogen activator inhibitor-1 and glomerulosclerosis in vivo. Kidney Int. 58, 1219–1227 (2000).

    Article  CAS  Google Scholar 

  25. Niranjan, T. et al. The Notch pathway in podocytes plays a role in the development of glomerular disease. Nat. Med. 14, 290–298 (2008).

    Article  CAS  Google Scholar 

  26. Isermann, B. et al. Activated protein C protects against diabetic nephropathy by inhibiting endothelial and podocyte apoptosis. Nat. Med. 13, 1349–1358 (2007).

    Article  CAS  Google Scholar 

  27. Pitt, B. & Leopold, J.A. A role for mineralocorticoid receptor blockade in the prevention of apoptosis. J. Mol. Cell. Cardiol. 39, 415–417 (2005).

    Article  CAS  Google Scholar 

  28. Wu, X. et al. Rac1 activation controls nuclear localization ofβ-catenin during canonical Wnt signaling. Cell 133, 340–353 (2008).

    Article  CAS  Google Scholar 

  29. Liu, W.F., Nelson, C.M., Tan, J.L. & Chen, C.S. Cadherins, RhoA and Rac1 are differentially required for stretch-mediated proliferation in endothelial versus smooth muscle cells. Circ. Res. 101, e44–e52 (2007).

    Article  CAS  Google Scholar 

  30. Takenaka, T., Forster, H., De Micheli, A. & Epstein, M. Impaired myogenic responsiveness of renal microvessels in Dahl salt-sensitive rats. Circ. Res. 71, 471–480 (1992).

    Article  CAS  Google Scholar 

  31. Eto, N. et al. Podocyte protection by darbepoetin: preservation of the cytoskeleton and nephrin expression. Kidney Int. 72, 455–463 (2007).

    Article  CAS  Google Scholar 

  32. Komuro, R., Sasaki, T., Takaishi, K., Orita, S. & Takai, Y. Involvement of Rho and Rac small G proteins and Rho GDI in Ca2+-dependent exocytosis from PC12 cells. Genes Cells 1, 943–951 (1996).

    Article  CAS  Google Scholar 

  33. Diekmann, D. et al. Bcr encodes a GTPase-activating protein for p21rac. Nature 351, 400–402 (1991).

    Article  CAS  Google Scholar 

  34. Iida, T. et al. Functional modulation of the mineralocorticoid receptor by cis-diamminedichloroplatinum (II). Kidney Int. 58, 1450–1460 (2000).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to S. Fukuda for help in electron microscopic analysis. We thank Pfizer for providing eplerenone and Asahi Kasei Pharma for providing fasudil.

Author information

Author notes

  1. Shigeru Shibata and Miki Nagase: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Nephrology and Endocrinology, University of Tokyo Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Shigeru Shibata, Miki Nagase, Shigetaka Yoshida, Wakako Kawarazaki & Toshiro Fujita

  2. Department of Anatomy, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan

    Hidetake Kurihara

  3. Division of Clinical Immunology, Advanced Clinical Research Center, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan

    Hirotoshi Tanaka

  4. Department of Molecular Biology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka, 537-8511, Japan

    Jun Miyoshi

  5. Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan

    Yoshimi Takai

Authors
  1. Shigeru Shibata
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Miki Nagase
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Shigetaka Yoshida
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Wakako Kawarazaki
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Hidetake Kurihara
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Hirotoshi Tanaka
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Jun Miyoshi
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Yoshimi Takai
    View author publications

    You can also search for this author inPubMed Google Scholar

  9. Toshiro Fujita
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

S.S. and M.N. planned and performed experiments and wrote the manuscript. S.Y. and W.K. helped with experimental procedures and contributed to data discussion. H.K. provided the podocyte cell line and advised on the technical proposal. H.T. provided mineralocorticoid receptor plasmids and advised on the experimental approach and writing. J.M. and Y.T. generated Arhgdia−/− mice, provided expression plasmids containing wild-type and mutant Rho GTPases, advised on the experimental approach and contributed to data discussion. T.F. planned and directed the project and reviewed the manuscript.

Corresponding author

Correspondence to Toshiro Fujita.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–4 (PDF 2480 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shibata, S., Nagase, M., Yoshida, S. et al. Modification of mineralocorticoid receptor function by Rac1 GTPase: implication in proteinuric kidney disease. Nat Med 14, 1370–1376 (2008). https://doi.org/10.1038/nm.1879

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.1879

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing