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The nuclear interactome of 14-3-3zeta reveals its critical roles for genome accessibility during early stages of adipogenesis

ABSTRACT: Little is known about the molecular factors that tightly coordinate sequential actions of adipogenic cellular pathways that occur during the early stages of adipocyte precursor cells (APCs) differentiation. We previously established the scaffold protein, 14-3-3ζ, as a critical regulator of adipogenesis (Lim et al., Nat Commun., 2015). Our proteomic analysis performed in the present study suggested functional interactions between 14-3-3ζ and epigenetic effectors, such as DNMTs and HDACs, during the intial stages of APC differentiation. To assess if or how 14-3-3ζ influences chromatin accessibility in differentiating APCs, we performed ATAC-seq on 3T3-L1 preadipocytes that were transfected with a control, non-targeting siRNA (siCon) or siRNA against Ywhaz (sizeta), the gene encoding 14-3-3ζ (using either a Ywhaz-targeting or a control siRNA), followed by differentiation with the adipogenic cocktail (MDI) for 0h, 24h and 48h. Our ATAC-seq analysis revealed that 14-3-3ζ depletion impacted the accessibility of up to 1,244 chromatin regions corresponding in part to adipogenic genes, promoters, and enhancers during the initial 48 hours of differentiation. In conclusion, we report 14-3-3ζ as a key epigenetic modulator during induction of adipogenesis.

ORGANISM(S): Mus musculus

PROVIDER: GSE262016 | GEO | 2025/08/18

REPOSITORIES: GEO

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Project description:Background: Obesity is characterized as a disease that directly affects the whole-body metabolism and is associated with excess fat mass and several related comorbidities. Dynamics of the adipocyte hypertrophy and hyperplasia play an important role in health and disease, especially in obesity. Human adipose-derived stem cells (hASC) represent an important source for understanding the entire adipogenic differentiation process. However, little is known about the triggering step of adipogenesis in hASC. Here, we performed a proteogenomic approach for understanding the protein abundance alterations expression changes during the initiation of the adipogenic differentiation process. Methods: hASC were isolated from adipose tissue from three donors, characterized and expanded. Cells were cultured for 24 hours in adipogenic differentiation medium followed to protein extraction. We used shotgun proteomics to compare the proteomic profile of 24h-adipogenic differentiated and undifferentiated hASC. Besides, we used our previously next-generation sequencing data (RNA-seq) from the total and polysomal mRNA fractions of hASC to study post-transcriptional regulation during the initial steps of adipogenesis. Results: We identified a total of 3,.420 proteins out of and 48,.336 peptides, being 92 exclusively identified proteins in the undifferentiated hASC and 53 exclusive proteins in 24h-differentiated cells. Using a stringent criterion, we identified 33 differentially abundant proteins when compare 24h-differentiated versus undifferentiated hASC (14 upregulated and 19 downregulated, respectively). Among the upregulated proteins, we shortlist identified several adipogenic-related proteins. Combined analysis of the proteome and the transcriptome allowed the identification of positive correlation coefficients between proteins and mRNAs. Conclusions: These results demonstrate a specific proteome profile related to adipogenesis at the very beginning (24h) of the differentiation process in hASC, which represents an important piece for a better understanding of human adipogenesis and obesity. In addition, the adipogenic differentiation is fine-tuning regulated at transcriptional, post-transcriptional and post-translational levels.

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The nuclear interactome of 14-3-3zeta reveals its critical roles for genome accessibility during early stages of adipogenesis

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