Limits...
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

Gene expression analysis after TAF7L knockdown in C3H10T1/2 cells.(A)–(F) Time course of gene expression by RT-qPCR analysis of Taf7l (A), Pparγ (B), Adipoq (C), Glut4 (D), Fabp4 (E), and Klf15 (F) in C3H10T1/2 cells stably treated with shGFP or shTAF7L sequences at 0D, 1D, 3D, and 5D post adipogenic induction. D, days; shGFP, control cells; shTAF7L, TAF7L knockdown cells. mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D were assigned to 1, mRNA levels of each gene at 0D, 1D, 3D, and 5D in both shGFP and shTAF7L-treated C3H10T1/2 cells during adipogenesis were compared to mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D respectively, data is mean from triplicates.DOI:http://dx.doi.org/10.7554/eLife.00170.007
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3539393&req=5

fig2s1: Gene expression analysis after TAF7L knockdown in C3H10T1/2 cells.(A)–(F) Time course of gene expression by RT-qPCR analysis of Taf7l (A), Pparγ (B), Adipoq (C), Glut4 (D), Fabp4 (E), and Klf15 (F) in C3H10T1/2 cells stably treated with shGFP or shTAF7L sequences at 0D, 1D, 3D, and 5D post adipogenic induction. D, days; shGFP, control cells; shTAF7L, TAF7L knockdown cells. mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D were assigned to 1, mRNA levels of each gene at 0D, 1D, 3D, and 5D in both shGFP and shTAF7L-treated C3H10T1/2 cells during adipogenesis were compared to mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D respectively, data is mean from triplicates.DOI:http://dx.doi.org/10.7554/eLife.00170.007

Mentions: Next, we measured gene expression programs in differentiated C3H10T1/2 cells after depletion of TAF7L (10T1/2-shTAF7L-post). Importantly, genes normally up-regulated following adipogenesis (Figure 3A, shown in orange) showed similarly low expression levels in induced shTAF7L cells as in pre-adipocytes (Figure 3B, orange). Strikingly, 2083 out of 2360 genes (88%) that are highly upregulated during adipocyte differentiation failed to be induced upon adipogenic induction in the absence of TAF7L (Figure 3B,C). RT-qPCR analysis of several representative marker genes confirmed that TAF7L knockdown dramatically reduced their mRNA levels compared to control shGFP in post-adipogenesis cells (Figure 2C and Figure 2—figure supplement 1). These data suggest that the induction of adipocyte-specific genes is markedly compromised in the absence of TAF7L. Moreover, the overall transcriptional profile of differentiated shTAF7L-treated C3H10T1/2 cells (shTAF7L-post) largely matched the expression levels of pre-differentiated C3H10T1/2 cells (10T1/2-pre) (R = 0.98) (Figure 3B); while mature adipocytes (10T1/2-post) is distinct (R = 0.84) (Figure 3A). Thus, loss of TAF7L in C3H10T1/2 cells severely impaired its adipogenic potential rendering shTAF7L-treated cells in an undifferentiated state (Figures 2B and 3A,B). These findings were also confirmed using a different shTAF7L construct and RT-qPCR analysis (data not shown). Taken together, these results strongly implicate TAF7L in potentiating efficient adipogenesis of C3H10T1/2 cells by serving as an important regulator of adipocyte-specific gene expression.


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)

Gene expression analysis after TAF7L knockdown in C3H10T1/2 cells.(A)–(F) Time course of gene expression by RT-qPCR analysis of Taf7l (A), Pparγ (B), Adipoq (C), Glut4 (D), Fabp4 (E), and Klf15 (F) in C3H10T1/2 cells stably treated with shGFP or shTAF7L sequences at 0D, 1D, 3D, and 5D post adipogenic induction. D, days; shGFP, control cells; shTAF7L, TAF7L knockdown cells. mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D were assigned to 1, mRNA levels of each gene at 0D, 1D, 3D, and 5D in both shGFP and shTAF7L-treated C3H10T1/2 cells during adipogenesis were compared to mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D respectively, data is mean from triplicates.DOI:http://dx.doi.org/10.7554/eLife.00170.007
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3539393&req=5

fig2s1: Gene expression analysis after TAF7L knockdown in C3H10T1/2 cells.(A)–(F) Time course of gene expression by RT-qPCR analysis of Taf7l (A), Pparγ (B), Adipoq (C), Glut4 (D), Fabp4 (E), and Klf15 (F) in C3H10T1/2 cells stably treated with shGFP or shTAF7L sequences at 0D, 1D, 3D, and 5D post adipogenic induction. D, days; shGFP, control cells; shTAF7L, TAF7L knockdown cells. mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D were assigned to 1, mRNA levels of each gene at 0D, 1D, 3D, and 5D in both shGFP and shTAF7L-treated C3H10T1/2 cells during adipogenesis were compared to mRNA levels in shTAF7L-treated C3H10T1/2 cells at 0D respectively, data is mean from triplicates.DOI:http://dx.doi.org/10.7554/eLife.00170.007
Mentions: Next, we measured gene expression programs in differentiated C3H10T1/2 cells after depletion of TAF7L (10T1/2-shTAF7L-post). Importantly, genes normally up-regulated following adipogenesis (Figure 3A, shown in orange) showed similarly low expression levels in induced shTAF7L cells as in pre-adipocytes (Figure 3B, orange). Strikingly, 2083 out of 2360 genes (88%) that are highly upregulated during adipocyte differentiation failed to be induced upon adipogenic induction in the absence of TAF7L (Figure 3B,C). RT-qPCR analysis of several representative marker genes confirmed that TAF7L knockdown dramatically reduced their mRNA levels compared to control shGFP in post-adipogenesis cells (Figure 2C and Figure 2—figure supplement 1). These data suggest that the induction of adipocyte-specific genes is markedly compromised in the absence of TAF7L. Moreover, the overall transcriptional profile of differentiated shTAF7L-treated C3H10T1/2 cells (shTAF7L-post) largely matched the expression levels of pre-differentiated C3H10T1/2 cells (10T1/2-pre) (R = 0.98) (Figure 3B); while mature adipocytes (10T1/2-post) is distinct (R = 0.84) (Figure 3A). Thus, loss of TAF7L in C3H10T1/2 cells severely impaired its adipogenic potential rendering shTAF7L-treated cells in an undifferentiated state (Figures 2B and 3A,B). These findings were also confirmed using a different shTAF7L construct and RT-qPCR analysis (data not shown). Taken together, these results strongly implicate TAF7L in potentiating efficient adipogenesis of C3H10T1/2 cells by serving as an important regulator of adipocyte-specific gene expression.

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