人巨细胞病毒编码US28分子可促进CREB相关转录活性

Human cytomegalovirus-encoded US28 stimulates the CREB related transcriptional activity

目的:构建人巨细胞病毒(HCMV)编码的趋化因子受体类似物US28基因真核表达载体并观察其对通用转录因子磷酸化的环单磷酸腺苷反应元件结合蛋白(p-CREB)的表达量和相关转录活性的影响。方法:从病毒原液中扩增人巨细胞病毒编码US28基因,构建含US28基因真核表达载体,将该载体与CREB转录活性报告基因和恒定表达的内对照质粒组成的双萤光素酶报告基因系统共转染至HEK293细胞中,Western blot法检测CREB活性形式p-CREB的表达量,并分析US28促使的CREB相关的相对转录活性变化。结果:经PCR鉴定及测序结果证实,所构建的重组载体正确。US28可有效促进p-CREB的表达量与其提升转录的功能活性,揭示了US28与CREB相关转录活性增强的密不可分的关系。结论:人巨细胞病毒可利用宿主通用转录因子CREB增强相关基因转录,从而有可能促进病毒的活化进程。

AIM： To observe the effect of the human cytomegalovirus（HCMV）-encoded chemokine receptor homolog US28 on the human transcription factor CREB related transcriptional activity. METHODS： The US28 gene was cloned from DNA of HCMV-infected fibroblast at 72 h post infection. The amplified gene fragment was subsequently cloned into pcDNA3.1 eukaryotic expression vector. The recombinant plasmid was selected and identified by sequence analysis. US28-pcDNA3.1 was added to the DuaI-Luciferase Reporter Assay System. The immunoreactive bands of phospho-CREB（p-CREB） and luminescence values were observed. RESULTS： The constructed recombinant vector was verified by PCR analysis and DNA sequencing. US28 enhanced the transcriptional efficiency of CRE driving gene via p-CREB. CONCLUSION： HCMV could enhance the transcriptional activity of CRE driving gene via p-CREB. CREB might be involved in the very early reprogramming of the host cell during virus activation.