Recognition of Regulatory and Pathogenic RNA by Muscleblind Zinc Fingers

肌盲锌指识别调节性和致病性 RNA

基本信息

批准号：
7924930
负责人：
PETER Edwin WRIGHT
金额：
$ 7.77万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
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：
2009-09-30 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7924930
关键词：
5&apos Splice Site Adipocytes Affinity Area Binding Biochemical Biological Process Blood Vessels Cardiac Complex DNA Development Disease Double-Stranded RNA Exons Foundations Gene Expression Genetic Recombination Genetic Transcription Genetic Translation Goals Human Indium Integrins Lead Maps Mediating Messenger RNA Methods Molecular Muscle Mutagenesis Myotonic Dystrophy Neurons Nuclear Nucleic Acids Pathogenesis Photoreceptors Physiological Play Post-Transcriptional Regulation Process Proteins RNA RNA Binding RNA Sequences RNA Splicing RNA-Protein Interaction Regulation Reporting Research Research Activity Role Site Solutions Specificity Structure Therapeutic Translations Troponin T Untranslated RNA Work Zinc Fingers base cellular development insight mRNA Precursor novel programs research study stem stoichiometry three dimensional structure treatment strategy

5&apos Splice Site Adipocytes Affinity Area Binding Biochemical Biological Process Blood Vessels Cardiac Complex DNA Development Disease Double-Stranded RNA Exons Foundations Gene Expression Genetic Recombination Genetic Transcription Genetic Translation Goals Human Indium Integrins Lead Maps Mediating Messenger RNA Methods Molecular Muscle Mutagenesis Myotonic Dystrophy Neurons Nuclear Nucleic Acids Pathogenesis Photoreceptors Physiological Play Post-Transcriptional Regulation Process Proteins RNA RNA Binding RNA Sequences RNA Splicing RNA-Protein Interaction Regulation Reporting Research Research Activity Role Site Solutions Specificity Structure Therapeutic Translations Troponin T Untranslated RNA Work Zinc Fingers base cellular development insight mRNA Precursor novel programs research study stem stoichiometry three dimensional structure treatment strategy

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项目摘要

DESCRIPTION (provided by applicant): The biological functions of DNA and RNA are dependent upon highly specific and complex interactions with proteins. Elucidation of the mechanisms by which proteins recognize and manipulate nucleic acids in such fundamental processes as transcription, translation, mRNA splicing, replication, and recombination remains an area of intense research activity. Despite recent progress, the enormous diversity of protein-nucleic acid interactions means that many questions of fundamental importance remain unanswered. Multi-dimensional NMR and biochemical methods will be used to investigate the mechanisms by which the CCCH zinc finger protein muscleblind (MBNL) recognizes specific regulatory sites in single stranded RNA and also binds with high affinity to pathogenic double stranded RNA hairpins. Muscleblind contains four CCCH zinc finger motifs that are absolutely required for its RNA binding activity. Vertebrate muscleblind functions as a developmentally programmed regulator of alternative pre-mRNA splicing required for regulation of terminal muscle differentiation; it is also involved in differentiation of photoreceptors, neurons, adipocytes, and blood vessels. Muscleblind plays a central role in type 1 and type 2 myotonic dystrophy, diseases that are associated with CUG and CCUG repeat expansions in untranslated messenger RNA. The expanded RNA repeats accumulate in nuclear foci that aberrantly sequester muscleblind and thereby disrupt its normal, developmentally regulated splicing activities. The interactions of the human muscleblind zinc fingers with single stranded RNA targets and with double stranded CUG and CCUG repeats will be mapped using NMR and biochemical methods. Protein and RNA mutagenesis will be performed to identify the interactions that stabilize the single stranded and CUG and CCUG repeat RNA complexes and determine binding specificity and affinity. Solution structures of complexes with single stranded and duplex RNA molecules will be determined to elucidate the molecular basis for muscleblind function in normal cellular development and in RNA-mediated pathogenesis. This research will provide novel insights into the mechanisms by which the highly abundant CCCH zinc finger motif interacts with and recognizes diverse RNA targets, and will form the foundations for a structure-based description of the role of CCCH zinc finger proteins in post-transcriptional regulation of gene expression and their role in RNA-mediated pathogenesis.

描述（由申请人提供）：DNA和RNA的生物学功能取决于与蛋白质高度特异性和复杂的相互作用。阐明蛋白质在转录，翻译，mRNA剪接，复制和重组等基本过程中识别和操纵核酸的机制仍然是强烈的研究活动。尽管取得了最近的进步，但蛋白质核酸相互作用的巨大多样性意味着许多具有基本意义的问题仍未得到解答。多维的NMR和生化方法将用于研究CCCH锌指蛋白肌肉肌细胞（MBNL）识别单个绞合RNA中的特定调节位点的机制，并且还与致病性双束RNA RNA的高亲和力结合。 Muscleblind包含四个CCCH锌指基序，其RNA结合活性绝对需要。脊椎动物的肌肉闪光是调节末端肌肉分化所需的替代前MRNA剪接的开发调节剂；它还参与了光感受器，神经元，脂肪细胞和血管的分化。 Muscleblind在1型和2型肌发育症中起着核心作用，这些疾病与CUG和CCUG重复膨胀相关的疾病在未翻译的Messenger RNA中。扩展的RNA重复序列积累在核灶中，异常隔离了肌肉闪光，从而破坏了其正常的发育调节的剪接活性。人类肌肉闪烁锌指与单链RNA靶标的相互作用以及双链CUG和CCUG重复的相互作用将使用NMR和生化方法绘制。将进行蛋白质和RNA诱变，以确定稳定单个链，CUG和CCUG重复RNA复合物的相互作用，并确定结合特异性和亲和力。将确定具有单链和双链RNA分子的复合物的溶液结构，以阐明正常细胞发育和RNA介导的发病机理中肌肉闪线功能的分子基础。这项研究将提供有关高度丰富的CCCH锌指基序与不同的RNA靶标相互作用并识别的机制的新见解，并将为基础结构描述CCCH锌指蛋白在基因表达及其在RNA介导的病原体中的作用中的作用而形成基础。