College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Institute of Animal Husbandry, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China.
Biomolecules. 2023 Jun 29;13(7):1057. doi: 10.3390/biom13071057.
Long-chain acyl-CoA synthetase 1 (ACSL1) plays an important role in fatty acid metabolism and fat deposition. The transcription of the gene is regulated specifically among cells and physiological processes, and transcriptional regulation of in adipogenesis remains elusive. Here, we characterize transcription factors (TFs) associated with adipogenesis in the porcine gene. CCAAT-enhancer binding protein (C/EBP)α, a well-known adipogenic marker, was found to enhance the expression of the gene via binding two tandem motifs in the promoter. Further, we demonstrate that ACSL1 mediates C/EBPα effects on adipogenesis in preadipocytes cultured from subcutaneous fat tissue of pigs via gain- and loss-of-function analyses. The cAMP-response element binding protein, another TF involved in adipogenesis, was also identified in the regulation of gene expression. Additionally, single nucleotide polymorphisms (SNPs) were screened in the promoter of among four breeds including the Chinese indigenous Min, and Duroc, Berkshire, and Yorkshire pigs through sequencing of PCR products. Two tightly linked SNPs, -517G>T and -311T>G, were found exclusively in Min pigs. The haplotype mutation decreases promoter activity in PK-15 and ST cells, and in vivo the expression of , illustrating a possible role in adipogenesis regulated by C/EBPα/ACSL1 axis. Additionally, a total of 24 alternative splicing transcripts were identified, indicating the complexity of alternative splicing in the gene. The results will contribute to further revealing the regulatory mechanisms of during adipogenesis and to the characterization of molecular markers for selection of fat deposition in pigs.
长链酰基辅酶 A 合成酶 1（ACSL1）在脂肪酸代谢和脂肪沉积中发挥重要作用。该基因的转录在细胞和生理过程中受到特异性调节，而脂肪生成中 ACSL1 的转录调控仍不清楚。在这里，我们对猪 ACSL1 基因的脂肪生成相关转录因子（TFs）进行了特征描述。CCAAT 增强子结合蛋白（C/EBP）α是一种众所周知的脂肪生成标志物，通过结合启动子中的两个串联基序，发现其增强了 ACSL1 基因的表达。此外，我们通过对猪皮下脂肪组织来源的前体脂肪细胞进行功能获得和功能丧失分析，证明 ACSL1 介导了 C/EBPα 对脂肪生成的影响。参与脂肪生成的另一个 TF cAMP 反应元件结合蛋白也被鉴定为参与 ACSL1 基因表达的调控。此外，通过对包括中国本土民猪在内的四个品种（民猪、杜洛克猪、伯克夏猪和约克夏猪）的 PCR 产物测序，在 ACSL1 基因启动子中筛选出单核苷酸多态性（SNP）。发现两个紧密连锁的 SNP，-517G>T 和 -311T>G，仅存在于民猪中。单倍型突变降低了 PK-15 和 ST 细胞中的启动子活性，并且在体内，ACS1L 基因的表达降低，表明其可能在 C/EBPα/ACSL1 轴调控的脂肪生成中起作用。此外，还鉴定出 24 种可变剪接转录本，表明 ACSL1 基因在可变剪接方面的复杂性。这些结果将有助于进一步揭示 ACSL1 在脂肪生成过程中的调控机制，并为猪脂肪沉积的分子标记选择提供特征描述。
