HUMAN ISOFERRITINS STRUCTURE AND METABOLISM

人类异铁蛋白结构和代谢

• 批准号: 3225854
• 负责人: JAMES W DRYSDALE
• 金额: $ 19.35万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-07-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3225854
• 关键词: RNA splicing; cell free system; complementary DNA; electron microscopy; ferritin; gel electrophoresis; gene expression; genetic mapping; genetic regulation; genetic translation; genome; hereditary hemochromatosis; human tissue; in situ hybridization; iron metabolism; laboratory rabbit; laboratory rat; liver; messenger RNA; molecular cloning; neoplasm /cancer genetics; nucleic acid hybridization; nucleic acid sequence; protein biosynthesis; protein metabolism; protein structure; radiotracer; restriction mapping; ribonucleoproteins

This research will explore the organization, structure and expression of human ferritin genes. Cloned cDNA for H and L chains will be used to characterize the H and L gene families and to identify genes coding for the H and L subunits of tissue and serum ferritins. Their chromosomal location will be determined by in situ hybridization to chromosome spreads and by hybridization to DNA from separated chromosomes. Expressed genes will be identified from Southern analysis of DNA from rodent/human hybrid cell lines that express human H or L chains and by use of specific subprobes from cDNA clones and the 5' end of ferritin mRNAs. Genomic clones containing H and L genes will be isolated from genomic libraries. The organization and structure of expressed genes will be determined by use of restriction mapping and hybridization of the genomic clones to appropriate sub-probes, R loop and heteroduplex mapping, and finally by sequencing. Genes exhibiting conditional or tissue-specific differential expression will be identified and elements involved in possible regulation of their expression will be analyzed. Exon-intro junctions will be identified as well as possible regulatory sequences at the 5' end of the genes that may be involved in initiation or regulation of transcription or processing of tissue specific ferritins. These studies will eventually lead to an analysis of factors that might control the regulation of expression of H and L chains in different tissues and at different developmental stages. We shall investigate the possibility that defects in iron metabolism such as hemochromatosis or neonatal iron storage disease are causally related to abnormalities in ferritin expression. In related collaborative studies we shall explore the use of ferritin probes to determine whether any will be useful markers for identifying restriction fragment length polymorphisms associated with other inherited diseases where an altered gene lies close to ferritin. In parallel studies, we shall examine the interaction of ferritin mRNA and proteins and ribosomal RNA sequences in messenger ribonucleoprotein particles to explore their function in translational control of ferritin synthesis.

这项研究将探讨 人铁蛋白基因。 H和L链的克隆cDNA将用于 表征H和L基因家族，并确定编码的基因 组织和血清铁蛋白的H和L亚基。 他们的染色体位置 将通过原位杂交与染色体扩散和 分离的染色体与DNA杂交。 表达的基因将是 从啮齿动物/人类杂化细胞的DNA的南部分析中确定 表达人类H或L链并使用特定子探针的线条 来自cDNA克隆和铁蛋白mRNA的5'末端。 基因组克隆 包含H和L基因将从基因组文库中分离出来。 这 表达基因的组织和结构将通过使用 基因组克隆的限制映射和杂交 子探针，r环和异形图映射，最后是测序。 具有条件或组织特异性差异表达的基因 将被确定，并涉及可能调节其的要素 表达将进行分析。 外显子 - 内交叉将被确定为 在基因5'端的可能的调节序列尽可能 参与转录或处理的启动或调节 组织特异性铁蛋白。 这些研究最终将导致 分析可能控制h表达调节的因素 和不同组织和不同发育阶段的L链。 我们将调查铁代谢缺陷这样的可能性 由于血色素沉着病或新生儿铁储存疾病与 铁蛋白表达异常。 在相关的协作研究中我们 应探索使用铁蛋白探针来确定是否会 用于识别限制片段长度多态性的有用标记 与其他遗传性疾病相关的基因已改变的疾病关闭 到铁蛋白。 在并行研究中，我们将检查 铁蛋白mRNA和蛋白质和核糖体RNA序列 核糖核蛋白颗粒探索它们在翻译中的功能 铁蛋白合成的控制。