Abstract
Adipose tissue is the largest energy reservoir and secretory organ in the animal body, and is essential for maintaining normal physiological functions and metabolic balance. MicroRNAs regulate the process of adipogenic differentiation through post-transcriptional regulatory mechanisms. In the present study, miR-424 was upregulated during bovine adipocyte differentiation both in vivo and in vitro. The overexpression and interference of miR-424 exhibited the positive regulatory role in the differentiation of bovine adipocytes. Furthermore, miR-424 directly binds to the three prime untranslated region (3' UTR) of serine/threonine kinase 11 (STK11, also called LKB1), a master upstream gene in the AMP-activated protein kinase (AMPK) cascade, and up-regulates its expression. Functional studies showed that the knockdown of STK11 attenuated the pro-adipogenic effect of miR-424. Post-transcriptional regulation of STK11 by miR-424 was mediated potentially in an RNA binding protein (RBP) binding site-dependent manner. In conclusion, our study shows that miR-424 promotes bovine adipogenesis through an unconventional post-transcriptional regulation of STK11, which may serve as a potential target for the regulation of bovine adipogenesis and the improvement of livestock breeding efficiency.
Highlights
Adipose tissue is the main storage site for surplus energy, which is stored in the form of neutral triglycerides, and viewed as a biofuel reservoir that plays a crucial role in maintaining energy homeostasis (Sethi and Vidal-Puig, 2007)
MiR-424 appears to be involved in the regulation of bovine adipogenesis and was selected as the candidate miRNA for subsequent experiments
Reporter assays showed that the luciferase activity of the “F-FL” and “F-DMRE” reporters was significantly suppressed when miR-424 was absorbed by a specific inhibitor, but resulted in a modest yet significant increase in luciferase activity of the “F-miR-424 recognition element (MRE)” (Figure 5D). These results indicate that the presence of AU/GU rich sequences in the 3' UTR of STK11 might disrupt the miR-424/STK11 incorporation into the RNA-induced silencing complexes, suggesting that posttranscriptional regulation of STK11 by miR-424 occurs in a potential RNA binding protein (RBP) binding site-dependent manner
Summary
Adipose tissue is the main storage site for surplus energy, which is stored in the form of neutral triglycerides, and viewed as a biofuel reservoir that plays a crucial role in maintaining energy homeostasis (Sethi and Vidal-Puig, 2007). Adipocytes are the primary cell type in adipose tissue that deposit lipid droplets in response to overnutrition through the lipogenic pathway and release lipids during energy deficiency through the lipolytic pathway (Rutkowski et al, 2015). The intramuscular fat content in beef positively influences sensory quality traits and directly determines the meat grade (Hocquette et al, 2010; Park et al, 2018). Improving the fat content in beef is an essential goal of modern beef cattle breeding programs.
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