Red Cell Band 4.1 - Developmental Changes in RNA Splicing

红细胞带 4.1 - RNA 剪接的发育变化

基本信息

批准号：
7894777
负责人：
JOHN G CONBOY
金额：
$ 70.56万
依托单位：
UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7894777
关键词：
Actins Affinity Affinity Chromatography Alleles Alternative Splicing Animal Model Antibodies Binding Binding Sites Bioinformatics Biological Biological Assay Biological Models Biological Process Biology Brain Cell Line Cell membrane Cells Chimeric Proteins Coupled Data Defect Development Disease Distal Elements Enhancers Erythroblasts Erythrocytes Erythroid Erythroid Cells Event Exons Family Fishes Foxes Funding Genes Goals Human Individual Introns Investigation Lead Longitudinal Studies Mammals Maps Mass Spectrum Analysis Mechanics Mediating Membrane Modeling Molecular Mus Muscle Mutate Nature Nuclear Extract Play Protein Binding Protein Isoforms Proteins RNA RNA Splicing Regulation Regulatory Element Reporter Role Site Small Interfering RNA Specificity Spectrin Staging Structure System Technology Testing Therapeutic Time Tissue-Specific Splicing Tissues Trans-Splicing Western Blotting base cell type combinatorial comparative genomics erythroid differentiation expression vector genetic regulatory protein hnRNP A1 in vivo innovation insight interest mRNA Precursor mouse model novel paralogous gene programs protein activation research study spectrin-binding proteins vertebrate genome

项目摘要

This renewal proposal explores mechanisms that regulate alternative pre-mRNA splicing in differentiating erythroid cells, focusing on cis-regulatory sequences and trans-acting splicing factor proteins that regulate erythroid stage-specific switches in exon splicing. Studies will investigate the prototypical splicing switch in late erythroblasts that activates protein 4.1R exon 16 (E16), known to be physiologically important for mechanical stabilization of the red cell membrane, and several newly appreciated and evolutionarily conserved splicing switches in other erythroid pre-mRNAs. Major aims of the proposal include (1) affinity purification of novel factors that antagonize or synergize with Fox2 enhancer function in the highly conserved intron downstream of E16; (2) testing the hypothesis that four new erythroid splicing switches are coordinately regulated with E16, by Fox2 or other E16-regulatory factors, to provide new insights into the broader erythroid splicing program; and (3) initiation of a new effort to characterize functionality of cis-regulatory elements and trans-splicing factors in vivo with animal models. In addition to its biological importance for erythroid function, the E16 splicing switch is one of the best models for analysis of tissue-specific splicing in any cell system. Innovative features of the proposed studies include: detailed analysis of the conserved intron flanking the intron 16 Fox2 sites, that may provide insight into modulation of Fox-2 splicing activity; initial studies of the putatively co-regulated splicing switches in several other erythroid pre-mRNAs; and in vivo analysis of E16 regulatory elements. Successful accomplishment of these objectives will lead to a better understanding of the stage-specific switch in 4.1R premRNA splicing, and may provide preliminary evidence regarding a potential larger role for Fox-2 in mediating the erythroid differentiation stage-specific alternative splicing program. These studies should also provide insights into disease mechanisms caused by aberrant splicing, ultimately leading to splicing therapeutics to correct such defects.

这项更新提案探索了在分化红系细胞中调节替代前mRNA剪接的机制，重点关注调节外显子剪接中红系阶段特异性开关的顺式调节序列和反式作用剪接因子蛋白。研究将调查晚期成红细胞中激活蛋白 4.1R 外显子 16 (E16) 的原型剪接开关，已知该剪接开关对于红细胞膜的机械稳定性具有重要的生理意义，以及其他红细胞前体 mRNA 中新近认识到的和进化上保守的剪接开关。该提案的主要目标包括（1）亲和纯化E16下游高度保守的内含子中与Fox2增强子功能拮抗或协同的新因子； (2) 测试四个新的红细胞剪接开关与 E16、Fox2 或其他 E16 调节因子协调调节的假设，为更广泛的红细胞剪接程序提供新的见解； (3) 发起一项新的努力来表征顺式调控元件和反式剪接因子的功能体内动物模型。除了对红细胞功能具有生物学重要性外，E16 剪接开关还是分析任何细胞系统中组织特异性剪接的最佳模型之一。拟议研究的创新特征包括：对内含子 16 Fox2 位点侧翼的保守内含子进行详细分析，这可能有助于深入了解 Fox-2 剪接活性的调节；对其他几种红系前体 mRNA 中推定的共同调节剪接开关的初步研究；以及E16调控元件的体内分析。这些目标的成功实现将有助于更好地理解 4.1R premRNA 剪接中的阶段特异性转换，并可能提供有关 Fox-2 在介导红细胞分化阶段特异性选择性剪接程序中潜在更大作用的初步证据。这些研究还应该提供对异常剪接引起的疾病机制的见解，最终导致剪接疗法来纠正此类缺陷。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Alternative pre-mRNA splicing switches modulate gene expression in late erythropoiesis.

选择性前 mRNA 剪接开关调节晚期红细胞生成中的基因表达。

DOI：
10.1182/blood-2008-05-160325
发表时间：
2009-04-02
期刊：
Blood
影响因子：
20.3
作者：
Miki L. Yamamoto;Tyson A. Clark;S. Gee;Jeong;A. Schweitzer;A. Wickrema;J. Conboy
通讯作者：
J. Conboy

Human erythroid 5-aminolevulinate synthase. Gene structure and species-specific differences in alternative RNA splicing.

人红系 5-氨基乙酰丙酸合酶。

DOI：
发表时间：
1992-09-15
期刊：
影响因子：
0
作者：
Conboy, J G;Cox, T C;Bottomley, S S;Bawden, M J;May, B K
通讯作者：
May, B K

Cloning of mDEAH9, a putative RNA helicase and mammalian homologue of Saccharomyces cerevisiae splicing factor Prp43.

mDEAH9 的克隆，mDEAH9 是一种假定的 RNA 解旋酶，也是酿酒酵母剪接因子 Prp43 的哺乳动物同源物。

DOI：
发表时间：
1997-10-28
期刊：
影响因子：
11.1
作者：
Gee, S;Krauss, S W;Miller, E;Aoyagi, K;Arenas, J;Conboy, J G
通讯作者：
Conboy, J G

The splicing regulatory element, UGCAUG, is phylogenetically and spatially conserved in introns that flank tissue-specific alternative exons.

剪接调控元件 UGCAUG 在系统发育和空间上保守于组织特异性替代外显子两侧的内含子中。

DOI：
发表时间：
2005
期刊：
影响因子：
14.9
作者：
Minovitsky, Simon;Gee, Sherry L;Schokrpur, Shiruyeh;Dubchak, Inna;Conboy, John G
通讯作者：
Conboy, John G

A correlation with exon expression approach to identify cis-regulatory elements for tissue-specific alternative splicing.

与外显子表达方法的相关性，以确定组织特异性选择性剪接的顺式调控元件。

DOI：
发表时间：
2007
期刊：
影响因子：
14.9
作者：
Das, Debopriya;Clark, Tyson A;Schweitzer, Anthony;Yamamoto, Miki;Marr, Henry;Arribere, Josh;Minovitsky, Simon;Poliakov, Alexander;Dubchak, Inna;Blume, John E;Conboy, John G
通讯作者：
Conboy, John G