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Dok1 mediates high-fat diet–induced adipocyte hypertrophy and obesity through modulation of PPAR-γ phosphorylation

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Abstract

Insulin receptor substrate (IRS)-1 and IRS-2 have dominant roles in the action of insulin1, but other substrates of the insulin receptor kinase, such as Gab1, c-Cbl, SH2-B and APS, are also of physiological relevance2,3,4,5. Although the protein downstream of tyrosine kinases-1 (Dok1) is known to function as a multisite adapter molecule in insulin signaling6,7,8, its role in energy homeostasis has remained unclear. Here we show that Dok1 regulates adiposity. Expression of Dok1 in white adipose tissue was markedly increased in mice fed a high-fat diet, whereas adipocytes lacking this adapter were smaller and showed a reduced hypertrophic response to this dietary manipulation. Dok1-deficient mice were leaner and showed improved glucose tolerance and insulin sensitivity compared with wild-type mice. Embryonic fibroblasts from Dok1-deficient mice were impaired in adipogenic differentiation, and this defect was accompanied by an increased activity of the protein kinase ERK and a consequent increase in the phosphorylation of peroxisome proliferator–activated receptor (PPAR)-γ on Ser112. Mutation of this negative regulatory site for the transactivation activity of PPAR-γ blocked development of the lean phenotype caused by Dok1 ablation. These results indicate that Dok1 promotes adipocyte hypertrophy by counteracting the inhibitory effect of ERK on PPAR-γ and may thus confer predisposition to diet-induced obesity.

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Figure 1: Diet-induced upregulation of Dok1 expression in WAT and reduced adiposity of Dok1−/− mice.
Figure 2: Impaired adipogenesis in the absence of Dok1.
Figure 3: Improved glucose tolerance and increased systemic insulin sensitivity in Dok1−/− mice fed a high-fat diet.
Figure 4: Effect of ERK-dependent phosphorylation of PPAR-γ on Ser112 on Dok1 regulation of adiposity.

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Change history

  • 23 January 2008

    In the version of this article initially published online, the second sentence of the fifth paragraph of the main text referred to "Emr1 (also known as F4180).” This should read "Emr1 (also known as F4/80).” The error has been corrected for all versions of the article.

References

  1. White, M.F. Insulin signaling in health and disease. Science 302, 1710–1711 (2003).

    Article  CAS  Google Scholar 

  2. Minami, A. et al. Increased insulin sensitivity and hypoinsulinemia in APS knockout mice. Diabetes 52, 2657–2665 (2003).

    Article  CAS  Google Scholar 

  3. Duan, C., Yang, H., White, M.F. & Rui, L. Disruption of the SH2-B gene causes age-dependent insulin resistance and glucose intolerance. Mol. Cell. Biol. 24, 7435–7443 (2004).

    Article  CAS  Google Scholar 

  4. Molero, J.C. et al. c-Cbl–deficient mice have reduced adiposity, higher energy expenditure and improved peripheral insulin action. J. Clin. Invest. 114, 1326–1333 (2004).

    Article  CAS  Google Scholar 

  5. Bard-Chapeau, E.A. et al. Deletion of Gab1 in the liver leads to enhanced glucose tolerance and improved hepatic insulin action. Nat. Med. 11, 567–571 (2005).

    Article  CAS  Google Scholar 

  6. Hosomi, Y. et al. Characterization of a 60-kilodalton substrate of the insulin receptor kinase. J. Biol. Chem. 269, 11498–11502 (1994).

    CAS  PubMed  Google Scholar 

  7. Carpino, N. et al. p62dok: a constitutively tyrosine-phosphorylated, GAP-associated protein in chronic myelogenous leukemia progenitor cells. Cell 88, 197–204 (1997).

    Article  CAS  Google Scholar 

  8. Yamanashi, Y. & Baltimore, D. Identification of the Abl- and rasGAP-associated 62-kDa protein as a docking protein, Dok. Cell 88, 205–211 (1997).

    Article  CAS  Google Scholar 

  9. Yamanashi, Y. et al. Role of the rasGAP-associated docking protein p62(dok) in negative regulation of B cell receptor-mediated signaling. Genes Dev. 14, 11–16 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Di Cristofano, A. et al. p62(dok), a negative regulator of Ras and mitogen-activated protein kinase (MAPK) activity, opposes leukemogenesis by p210(bcr-abl). J. Exp. Med. 194, 275–284 (2001).

    Article  CAS  Google Scholar 

  11. Wick, M.J., Dong, L.Q., Hu, D., Langlais, P. & Liu, F. Insulin receptor–mediated p62dok tyrosine phosphorylation at residues 362 and 398 plays distinct roles for binding GTPase-activating protein and Nck and is essential for inhibiting insulin-stimulated activation of Ras and Akt. J. Biol. Chem. 276, 42843–42850 (2001).

    Article  CAS  Google Scholar 

  12. Noguchi, T. et al. Tyrosine phosphorylation of p62(Dok) induced by cell adhesion and insulin: possible role in cell migration. EMBO J. 18, 1748–1760 (1999).

    Article  CAS  Google Scholar 

  13. Hosooka, T. et al. Inhibition of the motility and growth of B16F10 mouse melanoma cells by dominant negative mutants of Dok-1. Mol. Cell. Biol. 21, 5437–5446 (2001).

    Article  CAS  Google Scholar 

  14. Rosen, E.D. & Spiegelman, B.M. Molecular regulation of adipogenesis. Annu. Rev. Cell Dev. Biol. 16, 145–171 (2000).

    Article  CAS  Google Scholar 

  15. Desvergne, B., Michalik, L. & Wahli, W. Transcriptional regulation of metabolism. Physiol. Rev. 86, 465–514 (2006).

    Article  CAS  Google Scholar 

  16. Shinohara, H. et al. Dok-1 and Dok-2 are negative regulators of lipopolysaccharide-induced signaling. J. Exp. Med. 201, 333–339 (2005).

    Article  CAS  Google Scholar 

  17. Kubota, N. et al. PPAR-γ mediates high-fat diet-induced adipocyte hypertrophy and insulin resistance. Mol. Cell 4, 597–609 (1999).

    Article  CAS  Google Scholar 

  18. Miles, P.D., Barak, Y., He, W., Evans, R.M. & Olefsky, J.M. Improved insulin-sensitivity in mice heterozygous for PPAR-γ deficiency. J. Clin. Invest. 105, 287–292 (2000).

    Article  CAS  Google Scholar 

  19. Koutnikova, H. et al. Compensation by the muscle limits the metabolic consequences of lipodystrophy in PPAR-γ hypomorphic mice. Proc. Natl. Acad. Sci. USA 100, 14457–14462 (2003).

    Article  CAS  Google Scholar 

  20. He, W. et al. Adipose-specific peroxisome proliferator–activated receptor γ knockout causes insulin resistance in fat and liver but not in muscle. Proc. Natl. Acad. Sci. USA 100, 15712–15717 (2003).

    Article  CAS  Google Scholar 

  21. Jones, J.R. et al. Deletion of PPAR-γ in adipose tissues of mice protects against high fat diet-induced obesity and insulin resistance. Proc. Natl. Acad. Sci. USA 102, 6207–6212 (2005).

    Article  CAS  Google Scholar 

  22. Hu, E., Kim, J.B., Sarraf, P. & Spiegelman, B.M. Inhibition of adipogenesis through MAP kinase-mediated phosphorylation of PPAR-γ. Science 274, 2100–2103 (1996).

    Article  CAS  Google Scholar 

  23. Adams, M., Reginato, M.J., Shao, D., Lazar, M.A. & Chatterjee, V.K. Transcriptional activation by peroxisome proliferator–activated receptor γ is inhibited by phosphorylation at a consensus mitogen-activated protein kinase site. J. Biol. Chem. 272, 5128–5132 (1997).

    Article  CAS  Google Scholar 

  24. Camp, H.S. & Tafuri, S.R. Regulation of peroxisome proliferator–activated receptor γ activity by mitogen-activated protein kinase. J. Biol. Chem. 272, 10811–10816 (1997).

    Article  CAS  Google Scholar 

  25. Shao, D. et al. Interdomain communication regulating ligand binding by PPAR-γ. Nature 396, 377–380 (1998).

    Article  CAS  Google Scholar 

  26. Rangwala, S.M. et al. Genetic modulation of PPAR-γ phosphorylation regulates insulin sensitivity. Dev. Cell 5, 657–663 (2003).

    Article  CAS  Google Scholar 

  27. Cock, T.A., Houten, S.M. & Auwerx, J. Peroxisome proliferator–activated receptor-γ: too much of a good thing causes harm. EMBO Rep. 5, 142–147 (2004).

    Article  CAS  Google Scholar 

  28. Bluher, M. et al. Adipose tissue selective insulin receptor knockout protects against obesity and obesity-related glucose intolerance. Dev. Cell 3, 25–38 (2002).

    Article  CAS  Google Scholar 

  29. Leitner, J.W., Kline, T., Carel, K., Goalstone, M. & Draznin, B. Hyperinsulinemia potentiates activation of p21Ras by growth factors. Endocrinology 138, 2211–2214 (1997).

    Article  CAS  Google Scholar 

  30. Sakai, T. et al. Skp2 controls adipocyte proliferation during the development of obesity. J. Biol. Chem. 282, 2038–2046 (2007).

    Article  CAS  Google Scholar 

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Acknowledgements

T.H. performed most of the experiments and data analysis and wrote the manuscript. T. Noguchi supervised the study and wrote the manuscript. K.K. performed hyperinsulinemic-euglycemic clamp studies. T. Nakamura and H.S. contributed to the measurement of adipocyte size and number. H.I. and W.O. contributed to the fractionation of adipose tissue. K. Tobimatsu, K. Takazawa and M.S. assisted with in vitro biochemical analyses. Y.M. and R.H. assisted with real-time quantitative RT-PCR analysis. T.Y. assisted with analysis of metabolic parameters. M.A.L. and Y.Y. contributed to the generation of mice used in this study and reviewed the manuscript. M.K. directed the project as a principal investigator and reviewed the manuscript.

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This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) to M.K. and T. Noguchi; a grant for the Twenty-first Century Center for Excellence Program 'Center of Excellence for Signal Transduction Disease: Diabetes Mellitus as Model' from MEXT to M.K.; a grant for the Cooperative Link of Unique Science and Technology for Economy Revitalization from MEXT to M.K.; a Grant-in-Aid for Creative Scientific Research to M.K.; and US National Institutes of Health grant DK49780 to M.A.L. We thank Takeda Pharmaceutical for providing pioglitazone, Y. Tamori (Kobe University) for providing antibodies to PPAR-γ and an adenovirus vector encoding mouse Pparg, T. Satoh (Kobe University) for a GST fusion protein containing the Ras binding domain of c-Raf-1, K. Nakao (RIKEN Center for Developmental Biology) for generating mouse embroyos by in vitro fertilization and S. Hama for technical assistance.

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Correspondence to Masato Kasuga.

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Supplementary Figs. 1–7, Supplementary Table 1 and Supplementary Methods (PDF 1014 kb)

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Hosooka, T., Noguchi, T., Kotani, K. et al. Dok1 mediates high-fat diet–induced adipocyte hypertrophy and obesity through modulation of PPAR-γ phosphorylation. Nat Med 14, 188–193 (2008). https://doi.org/10.1038/nm1706

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