NOVEL IRON/HEMIN RESPONSIVE GENES IN HEMATOPOIESIS

造血中新型铁/血红素反应基因

• 批准号: 2391511
• 负责人: DAVID H BOLDT
• 金额: $ 16.49万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2391511
• 关键词: acid phosphatase; antisense nucleic acid; cell cycle; cell differentiation; deferoxamine; ferriheme; flow cytometry; gene expression; genetic library; genetic regulation; genetic regulatory element; genetic transcription; hematopoiesis; human genetic material tag; human tissue; in situ hybridization; iron; messenger RNA; northern blottings; nucleic acid sequence; polymerase chain reaction; protein kinase C; reporter genes; tissue /cell culture; transferrin

Iron is a critical nutritional element essential for a great variety of important biological processes including cell growth and differentiation, electron transfer reactions, and oxygen transport, activation, and detoxification. Also, increasing recent attention has focused on the role of iron in regulating gene expression. Iron regulation of gene expression occurs in bacteria, plants, yeast, and mammals including humans. This function of iron may provide a basis for further understanding the iron requirements of cell proliferation and differentiation. Although iron- responsive post-transcriptional gene regulation has been well described, transcriptional regulation by iron has been less well studied. Our laboratory has recently delineated a novel iron-dependent system of transcriptional regulation. At least two eukaryotic genes, protein kinase C-beta and tartrate resistant acid phosphatase/uteroferrin, are transcriptionally active in the presence of Fe (III) presented as ferric transferrin (FeTF), whereas both are inhibited at the transcriptional level in the presence of exogenous hemin (ferric protoporphyrin IX). It is now proposed to build upon these observations to identify additional genes whose expression is regulated by iron or hemin and to examine the roles of the products of iron/hemin regulated genes in hematopoietic cell proliferation and differentiation. The following specific aims are proposed: (1) to study the roles of the hTRAP and hPKC-beta genes, prototype iron/hemin responsive genes, in hematopoiesis; (2) to test the possibility that other mammalian genes with candidate hemin regulatory motifs may also be regulated by iron and hemin; (3) to identify by PCR differential hybridization screening )PDHS) and/or differential display PCR (DDRT-PCR) new hematopoietic cell genes which are activated or suppressed by iron or hemin and to determine their roles in hematopoietic cell differentiation; (4) to express the cloned genes identified in Aim 3 in a bacterial expression system and to examine effects of the purified, recombinant proteins on hematopoietic cell differentiation. Identification of a group of genes regulated at the transcriptional level by iron and hemin and elucidation of the possible functions of these genes in hematopoiesis will provide a basis for an expanded understanding of iron and related compounds in cellular homeostasis and should open new avenues for investigation in iron research.

铁是各种各样的关键营养元素 重要的生物学过程，包括细胞生长和分化， 电子转移反应以及氧气传输，激活和 解毒。 而且，越来越多的关注集中在角色上 铁在调节基因表达中的铁。 铁调节基因表达 发生在包括人类在内的细菌，植物，酵母和哺乳动物中。 这 铁的功能可能为进一步理解铁提供了基础 细胞增殖和分化的要求。 虽然铁 - 响应迅速的转录后基因调控已得到很好的描述， 铁的转录调节的研究较少。 我们的 实验室最近描述了一种新型的铁依赖性系统 转录调节。 至少两个真核基因，蛋白激酶 C-β和防sttrate抗性酸性磷酸酶/子宫铁蛋白是 在作为铁的Fe（III）存在下转录活跃 转铁蛋白（FETF），而两者在转录水平上受到抑制 在外源性半蛋白（铁核IX）的情况下。 现在 提议建立这些观察结果以识别其他基因 其表达受铁或hemin调节，并检查 造血细胞中铁/血素调节基因的产物 增殖和分化。 以下具体目的是 提议：（1）研究HTRAP和HPKC-BETA基因的作用， 原型铁/hemin反应性基因，造血中的原型； （2）测试 其他具有候选Hemin调节的哺乳动物基因的可能性 基序也可以受铁和脱糖的调节。 （3）通过PCR识别 差异杂交筛选）PDHS）和/或差异显示PCR （DDRT-PCR）被激活或抑制的新造血细胞基因 通过铁或血素，并确定它们在造血细胞中的作用 分化； （4）在A中鉴定出在AIM 3中鉴定的克隆基因 细菌表达系统并检查纯化的效果 造血细胞分化的重组蛋白。 鉴别 铁和 血液和阐明这些基因在 造血将为铁扩展的理解提供基础 和相关的蜂窝稳态中的化合物，应打开新的途径 用于铁研究的调查。