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Locus-linked Regulatory Motifs of Globin Gene Switching

球蛋白基因转换的位点连锁调节基序

基本信息

批准号：
7654229
负责人：
KENNETH R PETERSON
金额：
$ 36.75万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-06-15 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7654229
关键词：
Adult Affect Affinity African American Binding Binding Sites Biological Assay Bone Marrow Cells Boxing Cell Line Cells Chemicals Chromatin Complementary DNA Complex Computer software Cooley&apos s anemia DNA DNA Binding DNase-I Footprinting Data Databases Deoxyribonuclease I Deoxyribonucleases Depth Development Dimerization Direct Repeats Disease Distal Erythroid Erythroid Cells Erythropoiesis Family Fluorescence-Activated Cell Sorting Funding Gene Activation Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Transcription Globin Goals Greek Green Fluorescent Proteins Hereditary Disease Human Laboratories Lead Link Mass Spectrum Analysis Mediating Messenger RNA Modality Molecular Molecular Weight Mus Mutagenesis Mutation Mutation Spectra Nuclear Extract Nucleotides Oligonucleotides Open Reading Frames Patients Peptides Phenotype Point Mutation Polymerase Chain Reaction Progress Reports Promoter Regions Protein Binding Proteins Proteome Regulatory Element Relative (related person)Reporter Repression Repressor Proteins Research Reverse Transcriptase Polymerase Chain Reaction Role Sickle Cell Anemia Site Staging Structure Switch Genes Technology Testing Thalassemia Therapeutic Tissues Trans-Activators Transcription Initiation Site Transgenic Mice Transgenic Organisms Vertebrates base beta Thalassemia chromatin immunoprecipitation design fetal gel mobility shift assay gene repression genome database human GATA1 protein in vivo man mouse model mutant novel promoter protein expression therapeutic target vector yeast two hybrid system

项目摘要

[Analysis of gene regulation in vertebrates has long benefited from studies initiated with the p-globin locus. Fundamental principles of developmental stage- and tissue-specific control of transcription originated with the study of this complexly regulated locus. The overall goal of my research has been to understand basic regulatory mechanisms governing human p-like globin gene switching, in this instance, autonomous yglobin gene silencing during the adult stage of definitive erythropoiesis. We recently identified a novel Ayglobin gene silencer motif and an associated represser complex that are linked to the first new HPFH point mutation to be described in over 10 years. This silencer is located at -570 relative to the mRNA CAP site in a GATA binding motif and repression is mediated by GATA-1 binding at this site. Mutation of this site in our (3-YAC transgenic mouse model resulted in a HPFH phenotype. This phenotype is corroborated by the discovery of HPFH in a family with the analogous mutation in the Gy-9'c-bin gene (Luo et al., 2004). The proposed study will test the hypothesis that y-globin gene silencing is mediated, in part, through GATA-1 recruitment of co-repressors. We will assess the function of the -570 silencer in vivo, characterize the GATA-1-Mi2 represser complex that binds at this site and identify its component proteins. Several HPFH mutations are associated with a loss of GATA-1 binding. We will test the universality of this mechanism in vivo. Finally, we will generate and identify novel represser binding sites within the Ay-globin gene promoter using a cell-based reporter assay to select for HPFH mutations. These aims will extend our studies identifying the role of various c/s-regulatory motifs from the previous funding period into functional studies that will discern molecular mechanisms of y-globin gene silencing by trans-acting factors comprising represser complexes. These proteins may be developed into potential therapeutic targets to increase HbF.] Sickle cell disease (SCO), p-thalassemias and Cooley's anemia are common genetic diseases that affect millions of people worldwide. SCO alone impacts one of 500 African Americans born each year. Understanding the molecular mechanisms controlling globin gene switching may aid in the development of targeted therapies or therapeutics to treat these diseases, particularly research aimed at turning on the fetal y-globin genes, which has been shown to be effective for the treatment of SCO.

[对脊椎动物基因调控的分析长期以来受益于p-珠蛋白的研究