CELLULAR FACTORS DETERMINING THE VARIANT BEHAIVOR OF HPV 11

决定 HPV 11 变异行为的细胞因素

• 批准号: 6359595
• 负责人: KAREN J AUBORN
• 金额: $ 40.85万
• 依托单位: LONG ISLAND JEWISH MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6359595
• 关键词: DNA binding protein; gel mobility shift assay; gene expression; genetic promoter element; genetic transcription; host organism interaction; human papillomavirus; human tissue; interleukin 6; keratinocyte; larynx neoplasms; latent virus infection; point mutation; polymerase chain reaction; reporter genes; tissue /cell culture; viral carcinogenesis; virus DNA; virus genetics; virus related neoplasm /cancer; virus replication

Replication of HPVs is of two types. Maintenance replication occurs in basal cells of epithelium, and amplified replication only occurs in superficial layers of papillomas. Stopping maintenance replication would cure infected persons of HPV. Stopping amplified replication would eliminate spread of HPV. The premise of this proposal is that HPV replication is controlled, at least in part, by transcription factors which program differentiation of epithelial cells. Transcription factors control replication by binding at or near sites of initiation, or regulate transcription of proteins needed for replication. The hypothesis to be tested is that transcription factors belonging to the cEBPs family of transcription factors are part of the complex machinery that determines how HPVs replicate. cEBPs direct differentiation. cEBPs are expressed in papillomas and bind to the regulatory region of HPV11. Interleukin 6, which elevates cEBPbeta and cEBP6, is expressed in papillomas and induces proteins in differentiating cells. Aims will: 1) Determine whether transcription and/or replication of HPV11 DNA can be modulated by dumbbell oligonucleotides to cEBP binding motifs. In vivo assays will use these nuclease resistant oligomers to compete binding of cEBPs to HPV11 DNA. 2) Determine whether HPV replication and/or transcription can be modulated by point mutations of cEBP binding sites in the HPV11 DNA. 3) Determine whether HPV11 transcription and /or replication respond to the levels of cEBPbeta. An expression vector for cEBPbeta or addition of interleukin 6 will be used to change cEBPbeta levels. 4) Identify proteins from undifferentiated and differentiated keratinocytes and papillomas, that interact with the putative cEBP recognition sites in HPV11.

HPV的复制是两种类型。维护复制发生在 上皮的基底细胞和放大复制仅发生在 乳头状瘤的浅层层。停止维护复制将 治愈受感染的HPV的人。停止放大复制将 消除HPV的扩散。该提议的前提是HPV 复制至少部分由转录因子控制 哪个程序区分上皮细胞。转录因子 通过在启动或附近结合或调节来控制复制 复制所需的蛋白质的转录。假设是 测试的是属于Cebps家族的转录因子 转录因子是确定复杂机制的一部分 HPV如何复制。 Cebps直接分化。 Cebps在 乳头状瘤并结合HPV11的调节区域。白介素6， 升高Cebpbeta和Cebp6的，以乳头状瘤表示并诱导 分化细胞中的蛋白质。 目标将：1）确定HPV11的转录和/或复制是否 DNA可以通过哑铃寡核苷酸对CEBP结合基序调节。 体内测定将使用这些抗核酸酶抗性低聚物进行竞争 Cebps与HPV11 DNA的结合。 2）确定是否复制和/或 转录可以通过CEBP结合位点的点突变调节 HPV11 DNA。 3）确定HPV11是否转录和 /或 复制响应Cebpbeta的水平。表达向量的 Cebpbeta或添加白介素6将用于更改Cebpbeta 水平。 4）鉴定来自未分化和分化的蛋白质 角质形成细胞和乳头状瘤，与推定的CEBP相互作用 HPV11中的识别站点。