School of Forensic Medicine, China Medical University, Shenyang 110122, China.
Genes (Basel). 2019 Oct 12;10(10):802. doi: 10.3390/genes10100802.
This study identified a transcription factor that might bind to the 5' regulatory region of the and explored the potential effect on 5-HT1A receptor expression. Based on JASPAR predictions, the binding of the transcription factor was demonstrated using the electrophoretic mobility shift assay (EMSA). Vectors over-expressing the transcription factor were co-transfected into HEK-293 and SK-N-SH cells with the recombinant pGL3 vector, and relative fluorescence intensity was measured to determine regulatory activity. Additionally, the qRT-PCR and Western blot were also used to identify whether the transcription factor modulated the endogenous expression of 5-HT1A receptor. The results suggest that the transcription factor CCAA/T enhancer binding protein beta (CEBPB) likely binds to the -1219 to -1209 bp (ATG+1) region of the . Two sequences located in the -722 to -372 bp and -119 to +99 bp were also identified. Although the effect of CEBPB on endogenous 5-HT1A receptor expression was not significant, it exhibited the strong inhibition on the relative fluorescence intensity and the mRNA level of . CEBPB inhibited the human expression by binding to the sequence -1219 - -1209 bp. This is useful and informative for ascertaining the regulation of 5-HT1A receptor and mental diseases.
本研究鉴定出一种可能与结合的转录因子，并探讨其对 5-HT1A 受体表达的潜在影响。基于 JASPAR 的预测，使用电泳迁移率变动分析（EMSA）证实了转录因子的结合。将过表达转录因子的载体与重组 pGL3 载体共转染至 HEK-293 和 SK-N-SH 细胞中，并测量相对荧光强度以确定调节活性。此外，还使用 qRT-PCR 和 Western blot 来确定转录因子是否调节内源性 5-HT1A 受体的表达。结果表明，转录因子 CCAA/T 增强子结合蛋白β（CEBPB）可能与结合。还鉴定出位于-722 至-372 bp 和-119 至+99 bp 的两个序列。虽然 CEBPB 对内源性 5-HT1A 受体表达的影响不显著，但它对相对荧光强度和的 mRNA 水平表现出强烈的抑制作用。CEBPB 通过与-1219 至-1209 bp 序列结合抑制人表达。这对于确定 5-HT1A 受体和精神疾病的调节具有重要意义。
