绝经后骨质疏松症肾阴虚证关联基因CLCF1启动子区相互作用转录因子的筛选及分析

Screening and analysis of interacting transcription factor in the promoter region of human cardiotrophin-like cytokine 1

目的检测、分析与CLCF1基因上游启动子区域相互作用的转录因子。方法采用DNA pulldown技术联合质谱鉴定的方法检测与CLCF1基因启动子区域相互作用的蛋白,对鉴定出的蛋白质进行生物信息学分析,筛选出50个转录因子,使用平行反应监测(parallel reaction monitoring,PRM)技术靶向验证转录因子的表达情况。结果试验组与对照组共筛选出251个差异蛋白,GO富集结果表明差异蛋白主要涉及核糖体生物发生、正向调控DNA模板转录等生物学过程,包含转录调节复合物、SWI/SNF超家族型复合物,具有与cAMP应答元件结合、ATP水解活性等分子功能。KEGG通路显示,差异蛋白主要参与人t细胞白血病病毒1型感染、线粒体自噬、甲状腺激素等信号通路。PRM验证发现MAFK、TFE3、MITF、JUNB、FOSL2这5个转录因子表达趋势与DNA pulldown检测结果一致。结论MAFK、TFE3、MITF、JUNB、FOSL2这5个转录因子可能与CLCF1基因启动子区相结合。

Objective To detect and analyze transcription factor interacting with the upstream promoter region of CLCF1 gene.Methods DNA pulldown and mass spectrometry were used to detect the proteins interacting with the promoter region of CLCF1 gene.The identified proteins were analyzed using bioinformatics.Fifty candidate transcription factors were screened out.Parallel reaction monitoring(PRM)was used to verify the expression of the transcription factors.Results Two hundred and fifty-one proteins were screened in the experimental group and the control group.GO enrichment analysis showed that they were mainly involved in ribosome biogenesis,positive regulation of DNA-templated transcription,transcription regulator complex,SWI/SNF superfamily-type complex,cAMP response element binding,and ATP hydrolysis activity.KEGG pathway showed that these differential proteins were mainly involved in human T-cell leukemia virus 1 infection,mitophagy,and thyroid hormone signaling pathway.PRM verified thtat the expression trends of MAFK,TFE3,MITF,JUNB,and FOSL2 were consistent with the DNA pulldown detection result.Conclusion MAFK,TFE3,MITF,JUNB,and FOSL2 may bind to the promoter region of CLCF1 gene.