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Dual functions of TAF7L in adipocyte differentiation.

Zhou H, Kaplan T, Li Y, Grubisic I, Zhang Z, Wang PJ, Eisen MB, Tjian R - Elife (2013)

Bottom Line: Depletion of TAF7L reduced adipocyte-specific gene expression, compromised adipocyte differentiation, and WAT development as well.Genome-wide mRNA-seq expression profiling and ChIP-seq binding studies confirmed that TAF7L is required for activating adipocyte-specific genes via a dual mechanism wherein it interacts with PPARγ at enhancers and TBP/Pol II at core promoters.In vitro binding studies confirmed that TAF7L forms complexes with both TBP and PPARγ.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology , Howard Hughes Medical Institute, University of California, Berkeley , Berkeley , United States ; Li Ka Shing Center For Biomedical and Health Sciences, CIRM Center of Excellence, University of California, Berkeley , Berkeley , United States.

ABSTRACT
The diverse transcriptional mechanisms governing cellular differentiation and development of mammalian tissue remains poorly understood. Here we report that TAF7L, a paralogue of TFIID subunit TAF7, is enriched in adipocytes and white fat tissue (WAT) in mouse. Depletion of TAF7L reduced adipocyte-specific gene expression, compromised adipocyte differentiation, and WAT development as well. Ectopic expression of TAF7L in myoblasts reprograms these muscle precursors into adipocytes upon induction. Genome-wide mRNA-seq expression profiling and ChIP-seq binding studies confirmed that TAF7L is required for activating adipocyte-specific genes via a dual mechanism wherein it interacts with PPARγ at enhancers and TBP/Pol II at core promoters. In vitro binding studies confirmed that TAF7L forms complexes with both TBP and PPARγ. These findings suggest that TAF7L plays an integral role in adipocyte gene expression by targeting enhancers as a cofactor for PPARγ and promoters as a component of the core transcriptional machinery.DOI:http://dx.doi.org/10.7554/eLife.00170.001.

No MeSH data available.


Related in: MedlinePlus

TAF7L is required for the expression of adipocyte-specific genes.(A) and (B), mRNA-seq data on gene expression of C3H10T1/2 cells pre- (horizontal axis) and post-adipogenesis (vertical axis) (A); mRNA-seq data on gene expression in C3H10T1/2 cells pre-adipogenesis (horizontal axis) and C3H10T1/2 treated with shTAF7L post-adipogenesis (vertical axis) (B). Orange dots in (A) mark genes upregulated during adipogenesis; blue dots in (A) mark genes unchanged or downregulated during adipogenesis. Circled genes were tested individually in RT-qPCR analysis. R indicates the correlation of the expression programs between two compared cells (10T1/2-post vs 10T1/2-pre in (A), 10T1/2-shTAF7L-post vs 10T1/2-pre in (B)). (C) TAF7L knockdown blocks the upregulation of the adipocyte-specific genes which occurs during normal adipogenesis, pink circle represents 2360 genes upregulated in 10T1/2-post by 10-fold (10×) from (A); orange circle represents 2226 genes unchanged in 10T1/2-shTAF7L-post (B) compared to (A), 2083 genes in the overlapping intersect region account for 88% of total upregulated 10× genes in (A). (D) List of gene ontology analysis hits showing a few typical adipocyte genes involved in fat cell differentiation and metabolic processes.DOI:http://dx.doi.org/10.7554/eLife.00170.008
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fig3: TAF7L is required for the expression of adipocyte-specific genes.(A) and (B), mRNA-seq data on gene expression of C3H10T1/2 cells pre- (horizontal axis) and post-adipogenesis (vertical axis) (A); mRNA-seq data on gene expression in C3H10T1/2 cells pre-adipogenesis (horizontal axis) and C3H10T1/2 treated with shTAF7L post-adipogenesis (vertical axis) (B). Orange dots in (A) mark genes upregulated during adipogenesis; blue dots in (A) mark genes unchanged or downregulated during adipogenesis. Circled genes were tested individually in RT-qPCR analysis. R indicates the correlation of the expression programs between two compared cells (10T1/2-post vs 10T1/2-pre in (A), 10T1/2-shTAF7L-post vs 10T1/2-pre in (B)). (C) TAF7L knockdown blocks the upregulation of the adipocyte-specific genes which occurs during normal adipogenesis, pink circle represents 2360 genes upregulated in 10T1/2-post by 10-fold (10×) from (A); orange circle represents 2226 genes unchanged in 10T1/2-shTAF7L-post (B) compared to (A), 2083 genes in the overlapping intersect region account for 88% of total upregulated 10× genes in (A). (D) List of gene ontology analysis hits showing a few typical adipocyte genes involved in fat cell differentiation and metabolic processes.DOI:http://dx.doi.org/10.7554/eLife.00170.008

Mentions: To identify the full range of genes regulated by TAF7L in adipocyte differentiation, we performed mRNA-seq to profile global gene expression patterns in C3H10T1/2 cells prior to (10T1/2-pre) and after adipogenesis (10T1/2-post) (Figure 3A). Next, we verified our mRNA-seq results by single gene RT-qPCR assays for a handful of well-characterized adipocyte-specific genes such as Fabp4, Glut4, Adipsin, Lpl, as well as control genes such as Mef2c and Frzb (data not shown); these results confirmed high concordance between the RT-qPCR assays and the genome-wide mRNA-seq data, although RT-qPCR generally gave 3- to 4-fold higher sensitivity compared to mRNA-seq. We also surveyed a set of 2360 genes upregulated by 10-fold or more in C3H10T1/2 cells post- vs pre-differentiation (Figure 3); this analysis identified nearly all the well characterized adipocyte-specific genes including a large proportion of genes involved in adipocyte development and function (Figure 3D).10.7554/eLife.00170.008Figure 3.TAF7L is required for the expression of adipocyte-specific genes.


Dual functions of TAF7L in adipocyte differentiation.

Zhou H, Kaplan T, Li Y, Grubisic I, Zhang Z, Wang PJ, Eisen MB, Tjian R - Elife (2013)

TAF7L is required for the expression of adipocyte-specific genes.(A) and (B), mRNA-seq data on gene expression of C3H10T1/2 cells pre- (horizontal axis) and post-adipogenesis (vertical axis) (A); mRNA-seq data on gene expression in C3H10T1/2 cells pre-adipogenesis (horizontal axis) and C3H10T1/2 treated with shTAF7L post-adipogenesis (vertical axis) (B). Orange dots in (A) mark genes upregulated during adipogenesis; blue dots in (A) mark genes unchanged or downregulated during adipogenesis. Circled genes were tested individually in RT-qPCR analysis. R indicates the correlation of the expression programs between two compared cells (10T1/2-post vs 10T1/2-pre in (A), 10T1/2-shTAF7L-post vs 10T1/2-pre in (B)). (C) TAF7L knockdown blocks the upregulation of the adipocyte-specific genes which occurs during normal adipogenesis, pink circle represents 2360 genes upregulated in 10T1/2-post by 10-fold (10×) from (A); orange circle represents 2226 genes unchanged in 10T1/2-shTAF7L-post (B) compared to (A), 2083 genes in the overlapping intersect region account for 88% of total upregulated 10× genes in (A). (D) List of gene ontology analysis hits showing a few typical adipocyte genes involved in fat cell differentiation and metabolic processes.DOI:http://dx.doi.org/10.7554/eLife.00170.008
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fig3: TAF7L is required for the expression of adipocyte-specific genes.(A) and (B), mRNA-seq data on gene expression of C3H10T1/2 cells pre- (horizontal axis) and post-adipogenesis (vertical axis) (A); mRNA-seq data on gene expression in C3H10T1/2 cells pre-adipogenesis (horizontal axis) and C3H10T1/2 treated with shTAF7L post-adipogenesis (vertical axis) (B). Orange dots in (A) mark genes upregulated during adipogenesis; blue dots in (A) mark genes unchanged or downregulated during adipogenesis. Circled genes were tested individually in RT-qPCR analysis. R indicates the correlation of the expression programs between two compared cells (10T1/2-post vs 10T1/2-pre in (A), 10T1/2-shTAF7L-post vs 10T1/2-pre in (B)). (C) TAF7L knockdown blocks the upregulation of the adipocyte-specific genes which occurs during normal adipogenesis, pink circle represents 2360 genes upregulated in 10T1/2-post by 10-fold (10×) from (A); orange circle represents 2226 genes unchanged in 10T1/2-shTAF7L-post (B) compared to (A), 2083 genes in the overlapping intersect region account for 88% of total upregulated 10× genes in (A). (D) List of gene ontology analysis hits showing a few typical adipocyte genes involved in fat cell differentiation and metabolic processes.DOI:http://dx.doi.org/10.7554/eLife.00170.008
Mentions: To identify the full range of genes regulated by TAF7L in adipocyte differentiation, we performed mRNA-seq to profile global gene expression patterns in C3H10T1/2 cells prior to (10T1/2-pre) and after adipogenesis (10T1/2-post) (Figure 3A). Next, we verified our mRNA-seq results by single gene RT-qPCR assays for a handful of well-characterized adipocyte-specific genes such as Fabp4, Glut4, Adipsin, Lpl, as well as control genes such as Mef2c and Frzb (data not shown); these results confirmed high concordance between the RT-qPCR assays and the genome-wide mRNA-seq data, although RT-qPCR generally gave 3- to 4-fold higher sensitivity compared to mRNA-seq. We also surveyed a set of 2360 genes upregulated by 10-fold or more in C3H10T1/2 cells post- vs pre-differentiation (Figure 3); this analysis identified nearly all the well characterized adipocyte-specific genes including a large proportion of genes involved in adipocyte development and function (Figure 3D).10.7554/eLife.00170.008Figure 3.TAF7L is required for the expression of adipocyte-specific genes.

Bottom Line: Depletion of TAF7L reduced adipocyte-specific gene expression, compromised adipocyte differentiation, and WAT development as well.Genome-wide mRNA-seq expression profiling and ChIP-seq binding studies confirmed that TAF7L is required for activating adipocyte-specific genes via a dual mechanism wherein it interacts with PPARγ at enhancers and TBP/Pol II at core promoters.In vitro binding studies confirmed that TAF7L forms complexes with both TBP and PPARγ.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology , Howard Hughes Medical Institute, University of California, Berkeley , Berkeley , United States ; Li Ka Shing Center For Biomedical and Health Sciences, CIRM Center of Excellence, University of California, Berkeley , Berkeley , United States.

ABSTRACT
The diverse transcriptional mechanisms governing cellular differentiation and development of mammalian tissue remains poorly understood. Here we report that TAF7L, a paralogue of TFIID subunit TAF7, is enriched in adipocytes and white fat tissue (WAT) in mouse. Depletion of TAF7L reduced adipocyte-specific gene expression, compromised adipocyte differentiation, and WAT development as well. Ectopic expression of TAF7L in myoblasts reprograms these muscle precursors into adipocytes upon induction. Genome-wide mRNA-seq expression profiling and ChIP-seq binding studies confirmed that TAF7L is required for activating adipocyte-specific genes via a dual mechanism wherein it interacts with PPARγ at enhancers and TBP/Pol II at core promoters. In vitro binding studies confirmed that TAF7L forms complexes with both TBP and PPARγ. These findings suggest that TAF7L plays an integral role in adipocyte gene expression by targeting enhancers as a cofactor for PPARγ and promoters as a component of the core transcriptional machinery.DOI:http://dx.doi.org/10.7554/eLife.00170.001.

No MeSH data available.


Related in: MedlinePlus