Cheng Jin, Qin Wen-Juan, Balsai Nyamsuren, Shang Xuan-Jian, Zhang Meng-Ting, Chen Hong-Quan
School of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
School of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China; Anhui Agricultural University International Immunization Center, Hefei, 230036, China.
Anim Reprod Sci. 2020 Jul;218:106506. doi: 10.1016/j.anireprosci.2020.106506. Epub 2020 May 19.
Activity of transcription factors affect synthesis of G-protein coupled receptor 54 (GPR54), an important factor in regulation of initiation of puberty. Expression of the GPR54 gene in cattle is associated with polymorphisms in the proximal regulatory region (PRR) of the GPR54 gene. Transcription resulting in production of GPR54 mRNA transcript occurs as a result of transcription factor (TF) interactions in the PRR. Polymorphisms in the PRR may be associated with extent of activity of these TFs. Folliculogenesis-specific BHLH TF (FIGLA), neurogenin 2 (NEUROG2), and early growth response 1 (EGR1) are important in modulation of ovarian follicle development and neurons synthesizing GnRH, thus, regulating biosynthesis of luteinizing hormone. The aim of this study, therefore, was to assess the transcription-activating potential of binding sites for FIGLA, NEUROG2, and EGR1 TFs in the GPR54 promoter of cattle. Two luciferase-based promoters, ATC and CCT, which contain three single nucleotide polymorphisms (SNPs), A/C-794, T/C-663, and C/T-601, in the GPR54 PRR, were analyzed to evaluate gene expression and activation of different promoters by FIGLA, NEUROG2, and EGR1. The FIGLA induced GPR54 transcription through the CCT, whereas NEUROG2 and EGR1 induced GPR54 transcription through the ATC promoter-binding site. The CCT-activating effects of FIGLA were greater (2.56-fold) than the ATC-activating effects (P < 0.05). The ATC-activating effects of NEUROG2 and EGR1 were markedly greater (12.91- and 8.41-fold; P < 0.01) than CCT-activating effects. The polymorphisms, CCT and ATC, of the cattle GPR54 affect the activity of transcription factors, therefore, have an important effect on production of GPR54 mRNA transcript.
转录因子的活性会影响G蛋白偶联受体54（GPR54）的合成，GPR54是青春期启动调节中的一个重要因子。牛体内GPR54基因的表达与GPR54基因近端调控区（PRR）的多态性相关。GPR54 mRNA转录本的产生是PRR中转录因子（TF）相互作用的结果。PRR中的多态性可能与这些TF的活性程度有关。卵泡生成特异性BHLH转录因子（FIGLA）、神经生成素2（NEUROG2）和早期生长反应1（EGR1）在调节卵巢卵泡发育和合成促性腺激素释放激素（GnRH）的神经元中起重要作用，从而调节促黄体生成素的生物合成。因此，本研究的目的是评估牛GPR54启动子中FIGLA、NEUROG2和EGR1转录因子结合位点的转录激活潜力。分析了两个基于荧光素酶的启动子ATC和CCT，它们在GPR54 PRR中包含三个单核苷酸多态性（SNP），即A/C-794、T/C-663和C/T-601，以评估FIGLA、NEUROG2和EGR1对不同启动子的基因表达和激活情况。FIGLA通过CCT诱导GPR54转录，而NEUROG2和EGR1通过ATC启动子结合位点诱导GPR54转录。FIGLA对CCT的激活作用（2.56倍）大于对ATC的激活作用（P<0.05）。NEUROG2和EGR1对ATC的激活作用（分别为12.91倍和8.41倍；P<0.01）明显大于对CCT的激活作用。牛GPR54的多态性CCT和ATC影响转录因子的活性，因此，对GPR54 mRNA转录本的产生有重要影响。
