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转录因子 CEBPB 通过与体外 5'调控区结合抑制人 的表达。

Transcription Factor CEBPB Inhibits the Expression of the Human by Binding to 5' Regulatory Region in Vitro.

机构信息

School of Forensic Medicine, China Medical University, Shenyang 110122, China.

出版信息

Genes (Basel). 2019 Oct 12;10(10):802. doi: 10.3390/genes10100802.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b76/6827163/a4a03565b75b/genes-10-00802-g001.jpg

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