基础医学与临床2024，Vol.44Issue(1)：37-42,6.DOI:10.16352/j.issn.1001-6325.2024.01.0037
ATF6调控生殖相关基因HSPA1L表达的分子机制
Molecular mechanism of ATF6 regulating the reproduction related gene HSPA1L
摘要
Abstract
Objective To explore the effect of endoplasmic reticulum stress activating transcription factor 6(ATF6)on the expression of reproduction related gene heat shock protein family A member 1 like(HSPA1L)and preliminari-ly clarify its regulatory molecular mechanism.Methods The ATF6 over-expression plasmid was transfected into HEK-293T cells and the over-expression efficiency was verified by RT-qPCR and Western blot.The transcriptome sequen-cing information of testis tissue of male ATF6 knockout mice was used to screen five reproduction related genes down-stream of ATF6.The dual luciferase reporter assay selected the downstream genes with high promoter activity and de-tected the effect of over-expression of ATF6 on the promoter activity of downstream genes.The possible binding sites of ATF6 and downstream gene promoters were predicted by gene-regulation.RT-qPCR and Western blot were used to detect the effect of over-expression of ATF6 on downstream gene expression in HEK-293T cells.Whether ATF6 binds to downstream gene promoters was determined by electrophoretic mobility shift assay(EMSA).Results The expres-sion of ATF6 mRNA(P＜0.001)and protein(P＜0.001 and P＜0.05)in HEK-293T cells was significantly increased after transfection.HSPA1L(P＜0.001 and P＜0.05),a reproductive related gene downstream of ATF6 was screened by transcriptome sequencing and dual luciferase reporter assay.ATF6 promoted the truncated promoter activity of HSPA1L(P＜0.001).After over-expression of ATF6,the expression of HSPA1L was significantly increased(P＜0.001).The differences were statistically significant.ATF6 protein could bind to the aagtcgtcac DNA sequence of HSPA1L promoter.Conclusions ATF6,a key molecule of endoplasmic reticulum stress,regulates the expression of reproduction related gene HSPA1L by binding to the promoter of HSPA1L.This result will lay a foundation for further research on the prevention or treatment of endoplasmic reticulum stress(ERS)related male infertility.关键词
活化转录因子6/热休克蛋白A1样蛋白/男性生殖/基因调控Key words
activating transcription factor 6/heat shock protein family A member 1 like/male fertility/gene regulation分类
生物学引用本文复制引用
汪媛媛,朱席琳,伍晓盼,刘英..ATF6调控生殖相关基因HSPA1L表达的分子机制[J].基础医学与临床,2024,44(1):37-42,6.基金项目
国家重点研发计划(2021YFC2700601) （2021YFC2700601）
