Letter | Published:

Initiation of myoblast to brown fat switch by a PRDM16–C/EBP-β transcriptional complex

Nature volume 460, pages 11541158 (27 August 2009) | Download Citation

Abstract

Brown adipose cells are specialized to dissipate chemical energy in the form of heat, as a physiological defence against cold and obesity1. PRDM16 (PR domain containing 16) is a 140 kDa zinc finger protein that robustly induces brown fat determination and differentiation2. Recent data suggests that brown fat cells arise in vivo from a Myf5-positive, myoblastic lineage by the action of PRDM16 (ref. 3); however, the molecular mechanisms responsible for this developmental switch is unclear. Here we show that PRDM16 forms a transcriptional complex with the active form of C/EBP-β (also known as LAP), acting as a critical molecular unit that controls the cell fate switch from myoblastic precursors to brown fat cells. Forced expression of PRDM16 and C/EBP-β is sufficient to induce a fully functional brown fat program in naive fibroblastic cells, including skin fibroblasts from mouse and man. Transplantation of fibroblasts expressing these two factors into mice gives rise to an ectopic fat pad with the morphological and biochemical characteristics of brown fat. Like endogenous brown fat, this synthetic brown fat tissue acts as a sink for glucose uptake, as determined by positron emission tomography with fluorodeoxyglucose. These data indicate that the PRDM16–C/EBP-β complex initiates brown fat formation from myoblastic precursors, and may provide opportunities for the development of new therapeutics for obesity and type-2 diabetes.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data has been deposited in the Gene Expression Omnibus (GEO) public database under accession GSE15895.

References

  1. 1.

    & Brown adipose tissue: function and physiological significance. Physiol. Rev. 84, 277–359 (2004)

  2. 2.

    , & Transcriptional control of brown adipocyte development and physiological function–of mice and men. Genes Dev. 23, 788–797 (2009)

  3. 3.

    et al. PRDM16 controls a brown fat/skeletal muscle switch. Nature 454, 961–967 (2008)

  4. 4.

    , & Unexpected evidence for active brown adipose tissue in adult humans. Am. J. Physiol. Endocrinol. Metab. 293, E444–E452 (2007)

  5. 5.

    et al. Identification and importance of brown adipose tissue in adult humans. N. Engl. J. Med. 360, 1509–1517 (2009)

  6. 6.

    et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009)

  7. 7.

    et al. Functional brown adipose tissue in healthy adults. N. Engl. J. Med. 360, 1518–1525 (2009)

  8. 8.

    et al. Retinoblastoma protein functions as a molecular switch determining white versus brown adipocyte differentiation. Proc. Natl Acad. Sci. USA 101, 4112–4117 (2004)

  9. 9.

    et al. Rb and p107 regulate preadipocyte differentiation into white versus brown fat through repression of PGC-1α. Cell Metab. 2, 283–295 (2005)

  10. 10.

    et al. Nuclear receptor corepressor RIP140 regulates fat accumulation. Proc. Natl Acad. Sci. USA 101, 8437–8442 (2004)

  11. 11.

    et al. FOXC2 is a winged helix gene that counteracts obesity, hypertriglyceridemia, and diet-induced insulin resistance. Cell 106, 563–573 (2001)

  12. 12.

    et al. Regulation of the brown and white fat gene programs through a PRDM16/CtBP transcriptional complex. Genes Dev. 22, 1397–1409 (2008)

  13. 13.

    et al. Transcriptional control of brown fat determination by PRDM16. Cell Metab. 6, 38–54 (2007)

  14. 14.

    , & Stimulation of adipogenesis in fibroblasts by PPARγ2, a lipid- activated transcription factor. Cell 79, 1147–1156 (1994)

  15. 15.

    & Fat and beyond: the diverse biology of PPARγ. Annu. Rev. Biochem. 77, 289–312 (2008)

  16. 16.

    et al. A novel gene, MEL1, mapped to 1p36.3 is highly homologous to the MDS1/EVI1 gene and is transcriptionally activated in t(1;3)(p36;q21)-positive leukemia cells. Blood 96, 3209–3214 (2000)

  17. 17.

    et al. Overexpression of sPRDM16 coupled with loss of p53 induces myeloid leukemias in mice. J. Clin. Invest. 117, 3696–3707 (2007)

  18. 18.

    et al. Optimization and use of peptide mass measurement accuracy in shotgun proteomics. Mol. Cell. Proteomics 5, 1326–1337 (2006)

  19. 19.

    , , & Conditional ectopic expression of C/EBPβ in NIH-3T3 cells induces PPARγ and stimulates adipogenesis. Genes Dev. 9, 2350–2363 (1995)

  20. 20.

    Transcriptional control of adipocyte formation. Cell Metab. 4, 263–273 (2006)

  21. 21.

    & A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell 67, 569–579 (1991)

  22. 22.

    et al. CCAAT/enhancer binding protein-β is a transcriptional regulator of peroxisome-proliferator-activated receptor-γ coactivator-1α in the regenerating liver. Mol. Endocrinol. 22, 1596–1605 (2008)

  23. 23.

    , , & Defective adipocyte differentiation in mice lacking the C/EBPβ and/or C/EBPδ gene. EMBO J. 16, 7432–7443 (1997)

  24. 24.

    et al. Defective thermoregulation, impaired lipid metabolism, but preserved adrenergic induction of gene expression in brown fat of mice lacking C/EBPβ. Biochem. J. 389, 47–56 (2005)

  25. 25.

    et al. Lymphoproliferative disorder and imbalanced T-helper response in C/EBPβ-deficient mice. EMBO J. 14, 1932–1941 (1995)

  26. 26.

    & Formation of normally differentiated subcutaneous fat pads by an established preadipose cell line. J. Cell. Physiol. 101, 169–171 (1979)

  27. 27.

    et al. Complementary action of the PGC-1 coactivators in mitochondrial biogenesis and brown fat differentiation. Cell Metab. 3, 333–341 (2006)

  28. 28.

    & Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J. Cell Biol. 17, 299–313 (1963)

  29. 29.

    et al. A two-stage, p16(INK4A)- and p53-dependent keratinocyte senescence mechanism that limits replicative potential independent of telomere status. Mol. Cell. Biol. 22, 5157–5172 (2002)

  30. 30.

    , , , & TRB3 blocks adipocyte differentiation through the inhibition of C/EBPβ transcriptional activity. Mol. Cell. Biol. 27, 6818–6831 (2007)

  31. 31.

    & Episomal vectors rapidly and stably produce high-titer recombinant retrovirus. Hum. Gene Ther. 7, 1405–1413 (1996)

  32. 32.

    , & Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nature Protocols 2, 1896–1906 (2007)

  33. 33.

    et al. The International Protein Index: an integrated database for proteomics experiments. Proteomics 4, 1985–1988 (2004)

  34. 34.

    & Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nat. Methods 4, 207–214 (2007)

  35. 35.

    et al. Expression monitoring by hybridization to high-density oligonucleotide arrays. Nature Biotechnol. 14, 1675–1680 (1996)

  36. 36.

    & Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc. Natl Acad. Sci. USA 98, 31–36 (2001)

Download references

Acknowledgements

We are grateful to S. R. Farmer, J. Rheinwald and P. F. Johnson for providing cells and other reagents, R. Gupta for his critical comments on the manuscript, and J. Y. Choi and E. Naseri for their assistance. S.K. is supported by AHA scientist development grant (0930125N). P.S. is supported by a National Institutes of Health (NIH) grant (DK081605). This work was supported by grants from the Picower Foundation and the NIH (DK31405) to B.M.S., NIH HG3456 and GM67945 to S.P.G., and NIH/NCRR shared instrumentation grant S10-RR-023010.

Author Contributions S.K. and B.M.S. conceived and designed the experiments. S.K., K.K. and E.L. performed the experiments. All of the authors analysed the data. S.K. and B.M.S. wrote the paper.

Author information

Affiliations

  1. Dana-Farber Cancer Institute,

    • Shingo Kajimura
    • , Patrick Seale
    •  & Bruce M. Spiegelman
  2. Department of Cell Biology, Harvard Medical School, 44 Binney Street, Boston, Massachusetts 02115, USA

    • Shingo Kajimura
    • , Patrick Seale
    • , Kazuishi Kubota
    • , Steven P. Gygi
    •  & Bruce M. Spiegelman
  3. Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, USA

    • Elaine Lunsford
    •  & John V. Frangioni

Authors

  1. Search for Shingo Kajimura in:

  2. Search for Patrick Seale in:

  3. Search for Kazuishi Kubota in:

  4. Search for Elaine Lunsford in:

  5. Search for John V. Frangioni in:

  6. Search for Steven P. Gygi in:

  7. Search for Bruce M. Spiegelman in:

Corresponding author

Correspondence to Bruce M. Spiegelman.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-12 with Legends and Supplementary Tables 1-2.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature08262

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.