ROLE OF PRE-S2 PROMOTER OF HBV STUDIED IN HEPATOCELLULAR CARCINOMA CELL LINE

HBV Pre-S2 启动子在肝癌细胞系中的作用研究

• 批准号: 3853589
• 负责人: E TABOR
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3853589
• 关键词: DNA footprinting; chemical binding; clone cells; genetic promoter element; hepatitis B virus group; molecular site; nucleic acid sequence; nucleoproteins; virus DNA

The S open reading frame of the hepatitis B virus (HBV) comprises preS1 region, preS2 region, and S structural gene region, which code the large middle and major surface antigen polypeptides, respectively, in accordance with translation from the three in-frame initiation codons. In order to investigate the most important sequence within the preS2 promoter region, plasmids were created with serial deletion mutants directing the expression of the chloramphenicol acetyl transferase gene and the transcriptional activities were analyzed by transfection assays in four human hepatocellular carcinoma or hepatoblastoma cell lines (HepG2, PLC/PRF/5, Hep3B, and 2.2.15). Deletion mutant analysis revealed that the promoter from sequence 2861 to 2887 had highest activity. Two binding sites for nuclear proteins were observed by DNase I footprinting assay but multiple shifted DNA bands were identified by gel retardation assay suggesting that there are multiple binding sites for nuclear protein in the promoter sequence between nucleotides 2920 and 2951.

乙肝病毒S开放阅读框包括PreS1 区、前S2区和S结构基因区，它们编码的是大的 中间和主要表面抗原多肽分别根据 从三个框内起始密码子翻译而来。为了 研究preS2启动子区域内最重要的序列， 用系列缺失突变体来指导表达，从而创建了质粒 氯霉素乙酰转移酶基因及其转录产物的研究 通过转基因试验分析了四个人的活性。 肝细胞癌或肝母细胞瘤细胞系(HepG2，PLC/PRF/5， Hep3B和2.2.15)。缺失突变分析表明，该启动子 序列2861-2887的酶活性最高。两个结合位点 通过DNase I足迹试验观察到核蛋白，但多个 凝胶阻滞法检测到了移位的DNA条带。 启动子中有多个核蛋白结合部位 核苷酸2920和2951之间的序列。