DEVELOPMENTAL CONTROL OF CYTOKERATIN EXPRESSION

细胞角蛋白表达的发育控制

• 批准号: 2090664
• 负责人: ROBERT GLEN OSHIMA
• 金额: $ 46.79万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-05-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2090664
• 关键词: DNA footprinting; breast neoplasms; cell transformation; chromatin; developmental genetics; early embryonic stage; gene expression; gene targeting; genetic manipulation; genetic regulation; genetically modified animals; intermediate filaments; keratin; laboratory mouse; melanoma; molecular genetics; neoplastic cell; nucleic acid sequence; oncogenes; site directed mutagenesis; transcription factor

Keratin 8 (K8) and keratin 18 (K18) are the first intermediate filament proteins to be expressed during murine development, and represent the evolutionary ancestors of the more specialized keratins. In humans, K8 and K18 are the predominant intermediate filaments of internal organs and carcinomas of breast, liver, and intestine. Recently we have shown that oncogenes which activate the ras signal transduction pathway stimulate K18 expression through regulatory elements recognized by members of the ets and AP1 transcription factor families. In addition, we have generated mice which carry a null allele for the mouse K8 gene and shown that this gene is vital to the development of the embryo. This proposal focuses on both the regulation and function of these intermediate filaments. The functions of these proteins during both normal development and in tumor cells will be investigated with the use of gene targeting technology to generate mice and cells which lack these proteins. In particular, the possible function of these proteins in the invasive motility of both breast and melanoma tumor cells will be tested. The regulation of the K18 gene will be investigated by identifying both the trans-acting transcription factors necessary for expression and the sequences important for directing the gene to a chromatin state, either permissive for expression or incompatible with expression in different adult transgenic mouse tissues. Furthermore, because Kl8 expression in transgenic mice is found to be independent of its site of integration, efficient and directly proportional to the number of integrated genes, we will identify the sequences which are able to confirm these properties on heterologous genes.

角蛋白8（K8）和角蛋白18（K18）是第一个中间细丝 在鼠发育过程中要表达的蛋白质，并代表 更专业的角蛋白的进化祖先。在人类中，K8和 K18是内部器官的主要中间细丝 乳腺癌，肝脏和肠道癌。最近我们已经证明 激活RAS信号转导途径的致癌基因刺激K18 通过ETS成员认可的调节元素表达 和AP1转录因子家族。此外，我们已经产生了小鼠 携带小鼠K8基因的无效等位基因，并表明该基因 对于胚胎的发展至关重要。该提议重点介绍 这些中间丝的调节和功能。功能 在正常发育和肿瘤细胞中，这些蛋白质中的这些蛋白质将 通过使用基因靶向技术来研究小鼠 和缺乏这些蛋白质的细胞。特别是可能的功能 这些蛋白质在乳房和黑色素瘤的侵入性运动中 将测试肿瘤细胞。 K18基因的调节将是 通过识别两个反式转录因子来研究 表达和序列对于指导 染色质状态的基因，要么允许表达或 与不同成年转基因小鼠组织中表达不兼容。 此外，由于发现转基因小鼠中的Kl8表达为 独立于其整合地点，高效和直接 与综合基因的数量成正比，我们将确定 能够在异源上确认这些属性的序列 基因。