The Role of UBTF in Undiagnosed Neurodevelopmental Disorders

UBTF 在未确诊的神经发育障碍中的作用

基本信息

批准号：
9096370
负责人：
RODERICK T HORI
金额：
$ 22.8万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-06 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9096370
关键词：
Address Adult Affect Alleles Animal Model Behavioral Binding Biochemical Biological Models Boxing Cell Cycle Cell Cycle Checkpoint Cell Cycle Progression Cell Cycle Regulation Cell Nucleolus Cell Nucleus Cell physiology Cells Cessation of life ChIP-seq Chromatin Chromatin Structure Cognitive Collaborations Complex DNA Binding DNA Binding Domain DNA Damage DNA Polymerase I DNA Polymerase II Data Databases Disease Dysphonia Dystonia Event Exhibits Face Future Gene Expression Gene Structure Genes Genetic Genetic Transcription Genetic Variation Genetic study Genome HMGB Proteins Intervention Knock-out Lead Link Messenger RNA Mission Modeling Molecular Morphology Mus Mutation Mutism Neurodevelopmental Disorder Organism Outcome Parents Pathogenesis Pathologic Persons Phenotype Process Protein Isoforms Public Health RNA Recombinant DNA Reporting Research Ribosomal RNA Role Secondary Dystonia Shapes Site Stuttering System TOR1A gene Therapeutic Transcriptional Activation Tweens Variant Voice Work base cdc Genes disease diagnosis disease phenotype exome sequencing gain of function mutation gene function genetic variant genomic tools induced pluripotent stem cell innovation insight mouse model mutant nervous system disorder neurobehavioral personalized medicine programs promoter public health relevance rRNA Genes response therapeutic development transcription factor translational study

项目摘要

DESCRIPTION (provided by applicant): Recently, the Undiagnosed Disease Network identified a possibly pathogenic allele, UBTF p.E210K (Glu210Lys) that is heterozygous, in a subject that exhibited dystonia, stuttering, progression to mutism, neu- rodevelopmental delay and a high-pitched voice. There is a fundamental gap in understanding the genetic, cellular and biochemical relationship between this allele and these pathogenic phenotypes. It is an important problem to explicitly identify (validate) the pathogenic allele and understand the mechanism that leads from the allele to the disease because that information justifies and shapes the direction of future studies such as development of therapeutic molecules. Upstream Binding Transcription Factor (UBTF; also called UBF) is a transcription factor that exists as two isoforms, UBTF1 and UBTF2. UBTF binds DNA using its HMGB boxes and Glu210 follows a conserved hydrophobic core of HMGB box 2 in both isoforms. While UBTF1 regulates rRNA transcription by RNA pol I, UBTF2 regulates mRNA transcription by RNA pol II. The observed pathologic phenotypes overlap with those reported for isolated and secondary dystonias, and associated neurodevelopmental disorders. When examined, there is evidence that UBTF binds near the promoter of genes associated with dystonia, cell cycle progression and response to DNA damage. Based on this information, there is a critical need to: 1) develop an animal model with the variant E210K UBTF allele and 2) characterize alterations in UBTF activity due to the E210K mutation. The objective of this application is to generate UBTF mouse models with etiologic and face validity and determine how the E210K mutation alters UBTF's functions. Our central hypothesis is that the UBTF E210K allele is a gain-of-function mutation. Based on this framework of data related to UBTF1/2, the following specific aims will be pursued: a) Characterize phenotypes associated with mouse models harboring variant UBTF alleles; and b) Determine the molecular effects of the E210K mutation of previously defined roles of UBTF1 in the nucleolus and UBTF2 on expression of dystonia- associated and cell-cycle genes. In the first aim, conditional UBTF-knockout and UBTF E210K mice will be developed. Without a mammalian model system, the genetic relationship between UBTF E210K and the undiagnosed disease cannot be validated. In the second aim, the effect of the E210K mutation on DNA binding, transcriptional activation of putative target promoters and nucleolar morphology will be examined. Insights into the functional consequence of the E210K mutation may facilitate a deeper understanding of UBTF and cellular processes such as rRNA transcription, cell-cycle control and neurodevelopmental processes. The approach is innovative because it represents a departure from the status quo and shifts focus to UBTF's ability to regulate transcription by RNA pol II and identifies a previously unknown role for UBTF in neurological disease. These contributions will be significant because 1) they would constitute a successful application of personalized medicine to diagnose disease mechanisms and 2) validate the prediction that UBTF E210K is a pathogenic allele. 1

描述（由适用提供）：最近，未诊断的疾病网络确定了可能的致病等位基因，UBTF P.E210K（GLU210LYS）是杂合子的，在暴露于肌张力障碍，结结巴巴的受试者中，向突变，Neu-Rodevepental延迟延迟和较高的声音。在理解该等位基因与这些致病表型之间的遗传，细胞和生化关系方面存在根本差距。明确识别（验证）病原等位基因并了解导致等位基因到疾病的机制是一个重要的问题，因为这些信息可以证明并塑造未来研究的方向，例如热分子的发展。上游结合转录因子（UBTF;也称为UBF）是一种转录因子，它作为两个同工型UBTF1和UBTF2存在。 UBTF使用其HMGB框结合DNA，而GLU210遵循两个同工型中HMGB盒2的保守疏水核心。虽然UBTF1通过RNA Pol I调节RRNA转录，但UBTF2通过RNA Pol II调节mRNA转录。观察到的病理表型与报道的分离和继发性肌张力蛋白酶以及相关的神经发育障碍的表型重叠。在检查时，有证据表明UBTF在与肌张力障碍相关的基因启动子附近结合，细胞周期进程和对DNA损伤的反应。基于这些信息，至关重要的是：1）开发具有E210K UBTF等位基因变异的动物模型，而2）则表征由于E210K突变而导致的UBTF活动变化。该应用的目的是生成具有病因和面部有效性的UBTF小鼠模型，并确定E210K突变如何改变UBTF的功能。我们的中心假设是UBTF E210K等位基因是功能收益突变。基于与UBTF1/2相关的数据框架，将追求以下特定目的：a）表征与具有变异UBTF等位基因的鼠标模型相关的表型； b）确定UBTF1先前定义的E210K突变在Nucleolus和Ubtf2中对肌张障碍和细胞周期基因表达的分子效应。在第一个目标中，将开发有条件的UBTF-KNOCKOUT和UBTF E210K小鼠。没有哺乳动物模型系统，无法验证UBTF E210K与非驻留疾病之间的遗传关系。在第二个目标中，将检查E210K突变对DNA结合的影响，将检查推定靶启动子和核仁形态的转录激活。对E210K突变功能后果的见解可能有助于更深入地了解UBTF和细胞过程，例如RRNA转录，细胞周期控制和神经发育过程。这种方法具有创新性，因为它代表了与现状的不同，并将重点转移到UBTF通过RNA Pol II调节转录的能力，并确定了UBTF在神经疾病中的先前未知作用。这些贡献将是重要的，因为1）它们将构成个性化医学来诊断疾病机制的成功应用，以及2）验证UBTF E210K是病原等位基因的预测。 1