STAT1和组蛋白乙酰化修饰调控鸡lncRNA-BMP4转录研究

doi:10.16431/j.cnki.1671-7236.2022.09.006

Abstract

Abstract: 【Objective】 The purpose of this study was to investigate the factors affecting the transcription of long chain noncoding RNA-bone morphogenetic protein 4(lncRNA-BMP4) promoters in chickens, and study the molecular mechanisms regulating the specific expression of lncRNA-BMP4.【Method】 Using the muscle genome in chickens as a template, the promoter region of lncRNA-BMP4 in chickens was amplified by PCR and cloned, the lncRNA-BMP4-EFGP vector was constructed, and the lncRNA-BMP4 promoter activity was qualitatively analyzed.The core region of the lncRNA-BMP4 promoter was screened by chromosome 5'-terminal deletion and double luciferase system detection.The potential transcriptional factors of the regulatory core region were predicted and analyzed by online tools, and the transcriptional factors that really affected lncRNA-BMP4 were screened by point mutation and dual luciferase system.The transcriptional regulation of DNA methylation and histone acetylation on lncRNA-BMP4 was verified by apparent modification.【Result】 The lncRNA-BMP4 promoter fragments(1 288 bp) were successfully amplified and ligated with a pEGFP-N1 vector.After transfection into chicken fibroblast cell line (DF-1), it showed fluorescent expression, indicating that lncRNA-BMP4 promoter had promotive activity.The results of chromosome 5'-terminal deletion and double luciferase system detection showed that the core promoter region was-832 to-651 bp.The transcription factors screened by Jaspar database analysis showed that the core regions were SOX17, CREB1 and STAT1.Double luciferase system showed that STAT1 could promote the activity of lncRNA-BMP4 core promoter region, DNA methylation inhibitor 5'-Azacd had no effect on lncRNA-BMP4 transcriptional activity, while histone acetylation inhibitor TSA extremely significantly increased its transcriptional activity (P<0.01).【Conclusion】 This study suggested that the transcriptional activity of lncRNA-BMP4 was positively regulated by STAT1 and histone acetylation, while DNA methylation did not affect its transcription.The results provided a theoretical basis for a detailed analysis of the function and molecular mechanism of lncRNA-BMP4.

Key words: chickens; lncRNA; promoter; STAT1; epigenetic modification

CLC Number:

Q753

GAO Xiaomin, ZHOU Shujian, CHEN Chen, JIN Jing, HU Cai, ZHANG Chen, ZUO Qisheng, ZHANG Yani, CHEN Guohong, LI Bichun. Regulation of STAT1 and Histone Acetylation Modification on lncRNA-BMP4 Transcription in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3321-3332.

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Regulation of STAT1 and Histone Acetylation Modification on lncRNA-BMP4 Transcription in Chickens

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