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| Status |
Public on Mar 30, 2020 |
| Title |
Insulin-Induced Serine 22 Phosphorylation of Retinoid X Receptor Alpha Is Dispensable For Adipogenesis in Brown Adipocytes |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Insulin action initiates a series of phosphorylation events regulating cellular differentiation, growth and metabolism. We have previously discovered, in a mass spectrometry-based phosphoproteomic study, that insulin/IGF-1 signaling induces phosphorylation of retinoid x receptor alpha (RXRα) at S22 in mouse brown pre-adipocytes. Here, we show that insulin induces the phosphorylation of RXRα at S22 in both brown precursor and mature adipocytes through a pathway involving ERK, downstream of IRS-1 and -2. We also found that RXRα S22 phosphorylation is promoted by insulin and upon re-feeding in brown adipose tissue in vivo, and that insulin-stimulated S22 phosphorylation of RXRα is dampened by diet-induced obesity. We used Rxra knockout cells re-expressing wild type (WT) or S22A non-phosphorylatable forms of RXRα to further characterize the role of S22 in brown adipocytes. Knockout of Rxra in brown pre-adipocytes resulted in decreased lipid accumulation and adipogenic gene expression during differentiation, and re-expression of RxraWT alleviated these effects. However, we observed no significant difference in cells re-expressing the RxraS22A mutant as compared with the cells re-expressing RxraWT. Furthermore, comparison of gene expression during adipogenesis in the WT and S22A re-expressing cells by RNA sequencing revealed similar transcriptomic profiles. Thus, our data propose a dispensable role for RXRα S22 phosphorylation in adipogenesis and transcription in differentiating brown pre-adipocytes.
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| Overall design |
72 samples. 2 genotypes (WT and S22A), 2 cell cultures (biological replicate). For each biological replicate 3 timepoints (D0, D2, D6), and 3 technical replicate.
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| Contributor(s) |
Ardenkjær-Larsen J, Rupar K, Sinkevičiūtė G, Petersen PS, Villarroel J, Lundh M, Barrès R, Rabiee A, Emanuelli B |
| Citation(s) |
32249683 |
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| Submission date |
Mar 29, 2020 |
| Last update date |
Jul 01, 2020 |
| Contact name |
Lars Roed Ingerslev |
| E-mail(s) |
ingerslev@sund.ku.dk
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| Organization name |
Copenhagen University
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| Department |
NNF Center for Basic Metabolic Research
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| Lab |
Integrative Physiology
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| Street address |
Blegdamsvej 3B
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| City |
Copenhagen |
| ZIP/Postal code |
2200 |
| Country |
Denmark |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (36)
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| Relations |
| BioProject |
PRJNA616109 |
| SRA |
SRP254453 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE147687_RAW.tar |
133.3 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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