Molecular Biology of Human Erythrocyte Alpha Spectrin

人红细胞α血影蛋白的分子生物学

• 批准号: 7918635
• 负责人: PATRICK G GALLAGHER
• 金额: $ 41.38万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7918635
• 关键词: Address; Affect; Anemia; Architecture; Base Sequence; Binding; Biochemical; Biological Assay; Blood; Blood Transfusion; Cells; Chromatin; Code; DNA; DNA-Protein Interaction; Data; Defect; Deoxyribonuclease I; Disease; Enhancers; Epigenetic Process; Erythrocyte Membrane; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; Gene Expression; Gene Expression Regulation; Gene Structure; Genes; Genomics; Goals; Hemolytic Anemia; Hereditary Elliptocytosis; Hereditary Spherocytosis; Histones; Human; Hybridization Array; In Vitro; Inherited; Insulator Elements; Knowledge; Link; Maps; Membrane; Membrane Protein Gene; Membrane Proteins; Messenger RNA; Molecular; Molecular Biology; Mutation; Nucleic Acid Regulatory Sequences; Operative Surgical Procedures; Pathogenesis; Patients; Positioning Attribute; Principal Investigator; Protein Binding; RNA Polymerase II; Regulation; Regulatory Element; Reporter Genes; Role; Sequence Analysis; Site; Spectrin; Structure; Techniques; Technology; Tissues; Transcript; Transgenic Mice; alpha Spectrin; chromatin immunoprecipitation; genetic regulatory protein; genome sequencing; in vivo; insight; membrane biogenesis; membrane skeleton; mutant; programs; protein expression; public health relevance; research study; transcription factor

DESCRIPTION (provided by applicant): The long-term goals of this proposal are to elucidate the molecular mechanisms involved in normal and abnormal expression of the protein 1-spectrin, a critical component of the erythrocyte membrane skeleton. Abnormalities of ?-spectrin are associated with inherited hemolytic anemia, which is sometimes severe. The first aim of this proposal is to identify and characterize the cis-sequences, trans-factors, and epigenetic state, including chromatin architecture, across the ?-spectrin gene locus that regulate its expression in erythroid and nonerythroid cells. These studies address the hypothesis that common regulatory signatures in ?-spectrin and other erythrocyte membrane protein genes control their tissue-specific expression. Integration of mRNA transcript composition, genomic organization, RNA polymerase II binding, transcription factor and regulatory protein binding, and histone architecture will provide detailed knowledge of erythrocyte membrane gene structure, function, and regulation and allow us to identify a common regulatory signature that controls expression in erythroid cells. The studies in this aim combine high throughput genomic technologies with functional studies of gene expression. Techniques to be utilized include chromatin immunoprecipitation experiments followed by array hybridization (ChIP-chip) or whole genome sequencing (ChIP-seq), high throughput DNase I hypersensitive site mapping, and functional studies of gene expression. Results obtained from these studies will allow study of the role of 1-spectrin in erythropoiesis, membrane biogenesis, and inherited erythrocyte disorders. The second aim of this proposal is the identification of mutations that perturb ?- spectrin spectrin gene regulation and/or expression in patients with spectrin-linked inherited hemolytic anemias and characterization of the effect of these mutations on ?-spectrin gene structure, function, and expression. These studies address the hypothesis that defects of ?-spectrin occur in regions of functional importance and their elucidation will provide important information about the structure, function, and regulation of the ?-spectrin gene in normal and mutant erythrocytes. Nucleotide sequence analysis of amplified patient genomic DNA will be performed to identify genetic defects in cases of inherited hemolytic anemia associated with qualitative and quantitative defects of ?-spectrin. Previously known and newly identified cis-regulatory elements in the ?- spectrin gene will be interrogated and characterized in functional studies of gene regulation. Together, results from these studies will provide important information on the structure, function, and regulation of spectrin in erythroid and nonerythroid cells and shed additional insight into the pathogenesis of spectrin-linked disorders of the erythrocyte. PUBLIC HEALTH RELEVANCE: Many people suffer from anemia, or low blood count, due to abnormalities in the membranes, or lining, of their red blood cells. Some people require surgery or blood transfusions to treat the anemia. This application studies the what, why, and how of abnormalities of the lining of the red blood cell that cause the anemia.

描述（由申请方提供）：本提案的长期目标是阐明蛋白1-血影蛋白（红细胞膜骨架的关键组分）正常和异常表达所涉及的分子机制。什么人？-血影蛋白与遗传性溶血性贫血有关，这种贫血有时很严重。本提案的第一个目的是确定和表征顺式序列，反式因子和表观遗传状态，包括染色质结构，在整个？血影蛋白基因座调节其在红系和非红系细胞中表达。这些研究解决了以下假设：血影蛋白和其它红细胞膜蛋白基因控制它们的组织特异性表达。mRNA转录组成，基因组组织，RNA聚合酶II结合，转录因子和调节蛋白结合，组蛋白结构的整合将提供红细胞膜基因结构，功能和调控的详细知识，并允许我们确定一个共同的监管签名，控制红细胞系细胞中的表达。在这个目标的研究结合联合收割机高通量基因组技术与基因表达的功能研究。待利用的技术包括染色质免疫沉淀实验，然后进行阵列杂交（ChIP芯片）或全基因组测序（ChIP-seq），高通量DNase I超敏位点定位和基因表达的功能研究。从这些研究中获得的结果将允许研究1-血影蛋白在红细胞生成、膜生物发生和遗传性红细胞疾病中的作用。该建议的第二个目的是鉴定干扰？血影蛋白血影蛋白基因调控和/或血影蛋白连锁遗传性溶血性贫血患者的表达以及这些突变对？血影蛋白基因结构、功能和表达。这些研究提出了一个假设，即？血影蛋白存在于功能重要的区域，它们的阐明将提供有关血影蛋白的结构、功能和调节的重要信息。正常和突变红细胞中血影蛋白基因。将对扩增的患者基因组DNA进行核苷酸序列分析，以确定遗传性溶血性贫血病例中与？-血影蛋白以前已知的和新发现的顺式调节元件在？-血影蛋白基因将在基因调控的功能研究中被询问和表征。总之，从这些研究的结果将提供重要的信息血影蛋白的结构，功能和调节红细胞和nonerythroid细胞和脱落的发病机制血影蛋白相关疾病的红细胞。 公共卫生关系：许多人患有贫血症，或低血细胞计数，由于异常的膜，或内衬，他们的红细胞。有些人需要手术或输血来治疗贫血。这个应用程序研究什么，为什么，以及如何异常的红细胞的衬里，导致贫血。