Transcriptional control of preadipocyte determination by Zfp423

Abstract

The worldwide epidemic of obesity has increased the urgency to develop a deeper understanding of physiological systems related to energy balance and energy storage, including the mechanisms controlling the development of fat cells (adipocytes). The differentiation of committed preadipocytes to adipocytes is controlled by PPARγ and several other transcription factors1, but the molecular basis for preadipocyte determination is not understood. Using a new method for the quantitative analysis of transcriptional components, we identified the zinc-finger protein Zfp423 as a factor enriched in preadipose versus non-preadipose fibroblasts. Ectopic expression of Zfp423 in non-adipogenic NIH 3T3 fibroblasts robustly activates expression of Pparg in undifferentiated cells and permits cells to undergo adipocyte differentiation under permissive conditions. Short hairpin RNA (shRNA)-mediated reduction of Zfp423 expression in 3T3-L1 cells blunts preadipocyte Pparg expression and diminishes the ability of these cells to differentiate. Furthermore, both brown and white adipocyte differentiation is markedly impaired in Zfp423-deficient mouse embryos. Zfp423 regulates Pparg expression, in part, through amplification of the BMP signalling pathway, an effect dependent on the SMAD-binding capacity of Zfp423. This study identifies Zfp423 as a transcriptional regulator of preadipocyte determination.

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Figure 1: The C2H2 zinc-finger protein Zfp423 is enriched in preadipocytes.
Figure 2: Zfp423 regulates preadipocyte Pparg gene expression and adipocyte differentiation in vitro.
Figure 3: Zfp423 amplifies the pro-adipogenic actions of BMP proteins through its SMAD protein interaction domain.
Figure 4: Impaired brown and white adipocyte differentiation in Zfp423 -deficient embryos.

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Primary accessions

Gene Expression Omnibus

Data deposits

Complete microarray data is available at Gene Expression Omnibus under accession GSE19732 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE19732).

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Acknowledgements

We are grateful to S. Kleiner and S. Kajimura for critical reading of the manuscript and to all members of the Spiegelman laboratory for useful discussions. We thank B. Wagner for technical assistance in using robotic liquid handlers, B. Seed’s laboratory for help with high-through query of Primer bank, and J. Brestelli for performing high-throughput queries of the Primer 3 program. We are also grateful to D. Bernlohr for the FABP4 antiserum. R.K.G. is supported by the Ruth Kirstein NRSA (F32 DK079507-01), Z.A. is supported by K08 HL79172-01 (NHLBI) and the Smith Family Foundation Grant, P.S. is supported by NIH DK081605, and the research described in this study was supported by NIH DK31405 to B.M.S. and by NIDCD R01DC008295 to R.R.R.

Author Contributions R.K.G. and B.M.S. conceived and designed the experiments. R.K.G., Z.A., P.S., R.J.M., L.Y. and H.M.C. performed experiments. All authors analysed the data. Y.A.R., H.K. and R.R.R. provided reagents and samples, and R.K.G. and B.M.S. wrote the manuscript.

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Correspondence to Bruce M. Spiegelman.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-14 with legends, and Supplementary Tables 1-2. (PDF 1177 kb)

Supplementary Data

Supplementary Data Set 1: Raw Cycle Threshold data from Quanttrx assays (XLS 479 kb)

Supplementary Data

Supplementary Data Set 2: Primer sequences used in the Quanttrx Assays (XLS 522 kb)

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Gupta, R., Arany, Z., Seale, P. et al. Transcriptional control of preadipocyte determination by Zfp423. Nature 464, 619–623 (2010). https://doi.org/10.1038/nature08816

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