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Role of ZIC and GLI Protein-protein Interactions in Human Brain Disorders

ZIC 和 GLI 蛋白质-蛋白质相互作用在人脑疾病中的作用

基本信息

批准号：
7456781
负责人：
GERALD J WYCKOFF
金额：
$ 22.11万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-15 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7456781
关键词：
Brain Brain Diseases Calorimetry Cerebellum Chicago Child Class Clinical Collaborations Complex DNA Dandy-Walker Syndrome Data Development Diagnostic Disease Disruption Early Intervention Embryo Etiology Family Fingers Foundations Future GLI Family Protein GLI gene Generations Genes Genetic Goals Holoprosencephaly Human Human Development Individual Lead Life Life Expectancy Link Literature Methods Modeling Molecular Mutation Nature Numbers Patients Pattern Phase Phenotype Play Point Mutation Process Protein Family Proteins Proteomics Public Health Research Role Testing Thinking Time Titrations Two-Hybrid System Techniques Universities Walker-Warburg syndrome Walkers Work Yeasts Zinc Fingers brain malformation congenital brain disorder design developmental disease experience expression vector member mouse model mutant prognostic protein protein interaction theories tool transcription factor vector yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Mutations in Zinc Finger in the Cerebellum (ZIC) genes and the Glioma-associated Oncogene Homolog (GLI) have been linked to multiple debilitating, congenital brain disorders in humans. In particular, abnormalities in these proteins play a critical role in patterning disorders such as the Dandy-Walker Malformation, holoprosencephaly, and Greig Cephalopolysyndactyly Syndrome, with ramifications both for intellectual capacity and life expectancy. The implication of these genes in disease states has been proven in a variety of ways in the literature. For example, viable mouse models for Dandy-Walker are created through a mutation that fuses ZIC1and ZIC4, deleting zinc-finger domains from each. The precise mutations that cause these phenotypes in humans, however, have remained elusive. Attempts to examine Dandy-Walker and other patients have failed to uncover de novo mutations in ZIC and GLI proteins in a simple, causal relationship to the disease state. In other cases, substantial heterogenity exists between observed phenotypes even when mutations that cause a given disorder are present. We theorize that both of these issues arise from an incomplete understanding of ZIC functionality and interaction patterns. Until recently, the canonical model of zinc-finger proteins such as ZIC and GLI was that individual fingers in the protein never interacted with one another, but only with DNA. Recent research, however, has established that ZIC and GLI proteins likely do not function according to canonical models. Instead, they are members of a newly identified class of Cys2His2 proteins called two-finger or interacting-finger proteins, a subset characterized by interactions between fingers in a single protein. Furthermore, it has recently been proven that ZIC and GLI interact not only internally, from finger to finger, but with each other, in protein- protein interactions that remain understudied. These new observations, taken collectively, lead us to hypothesize that the disruption of ZIC-ZIC and ZIC-GLI interactions by mutations, and not single or paired mutations alone, are causal in the patterning disorders associated with these genes, particularly in Dandy-Walker Syndrome. We therefore propose a two-year study in which we will test whether interactions exist between ZIC proteins via yeast two-hybrid screens, confirm ZIC-ZIC and ZIC-GLI interactions via Isothermal Titration Calorimetry (ITC), and determine if mutations associated with disease states disrupt interactions between ZIC-ZIC and ZIC- GLI. Our hope is that the work will lead not only to an understanding of the molecular mechanisms underlying disorders associated with ZIC and GLI genes, but also to better prognostic and diagnostic clinical tests that will create opportunities for early intervention and lead to healthier and longer lives for children with some of these disorders. PUBLIC HEALTH RELEVANCE: This project will determine the role protein-protein interactions play in a set of devastating congenital brain disorders that manifest during human development. Guided by our recent findings on a class of zinc-finger proteins with a unique interaction domain, we will examine how mutation in these proteins causes or contributes to disease states, laying the foundation for future prognostic and diagnostic tools that will permit early intervention in developmental patterning disorders such as the Dandy-Walker Malformation, holoprosenchephaly, Grieg Cephalopolysyndactyly Syndrome, and others.

描述（由申请人提供）：小脑锌指（ZIC）基因和胶质瘤相关癌基因同源物（GLI）突变与人类多种衰弱性先天性脑部疾病有关。特别是，这些蛋白质的异常在模式化疾病中起着关键作用，如Dandy-Walker畸形，前脑无裂畸形和Greig头多指并指综合征，对智力和预期寿命都有影响。这些基因在疾病状态中的意义已经在文献中以各种方式得到证明。例如，Dandy-Walker的可行小鼠模型是通过融合ZIC 1和ZIC 4的突变创建的，从每个突变中删除锌指结构域。然而，导致人类这些表型的精确突变仍然难以捉摸。试图检查Dandy-Walker和其他患者未能发现ZIC和GLI蛋白的新生突变与疾病状态的简单因果关系。在其他情况下，即使存在导致给定病症的突变，观察到的表型之间也存在显著的异质性。我们的理论，这两个问题产生的ZIC功能和互动模式的不完整的理解。直到最近，锌指蛋白（如ZIC和GLI）的典型模型是蛋白质中的单个手指从不相互作用，而只与DNA相互作用。然而，最近的研究已经确定，ZIC和GLI蛋白可能不根据规范模型发挥作用。相反，它们是新发现的一类Cys 2 His 2蛋白质的成员，称为双指或相互作用指蛋白质，这是一个以单一蛋白质中手指之间的相互作用为特征的子集。此外，最近已经证明，ZIC和GLI不仅在内部相互作用，从手指到手指，而且在蛋白质-蛋白质相互作用中彼此相互作用，这仍然研究不足。这些新的观察结果，共同，使我们假设，ZIC-ZIC和ZIC-GLI相互作用的突变，而不是单一或成对的突变，是与这些基因相关的模式障碍，特别是在Dandy-Walker综合征的因果关系。因此，我们提出了一项为期两年的研究，其中我们将通过酵母双杂交筛选测试ZIC蛋白之间是否存在相互作用，通过等温滴定量热法（ITC）确认ZIC-ZIC和ZIC-GLI相互作用，并确定与疾病状态相关的突变是否破坏ZIC-ZIC和ZIC-GLI之间的相互作用。我们希望这项工作不仅能帮助我们了解与ZIC和GLI基因相关的疾病的分子机制，还能帮助我们更好地进行预后和诊断性临床测试，为早期干预创造机会，并为患有这些疾病的儿童带来更健康、更长寿的生活。公共卫生相关性：该项目将确定蛋白质-蛋白质相互作用在人类发育过程中表现出的一系列毁灭性先天性大脑疾病中的作用。通过我们最近对一类具有独特相互作用结构域的锌指蛋白的研究结果，我们将研究这些蛋白质中的突变如何导致或有助于疾病状态，为未来的预后和诊断工具奠定基础，这些工具将允许早期干预发育模式障碍，如Dandy-Walker畸形，全脑畸形，格里格头多指综合征等。