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.

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.

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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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