A multi-species approach to find regulators of deafness genes

寻找耳聋基因调节因子的多物种方法

基本信息

批准号：
10054189
负责人：
Andrew K Groves
金额：
$ 42.38万
依托单位：
BAYLOR COLLEGE OF MEDICINE
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：
2016-12-01 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10054189
关键词：
Adult Affect Amino Acids Ankle Auditory Biological Models Blindness Cell physiology Cells Data Development Disease Drosophila genus Etiology Future Genes Genetic Genetic Screening Genetic Variation Goals Grant Hair Cells Hearing Homologous Gene Human Insecta Invertebrates Label Labyrinth Link MYO7A gene Maintenance Mammalian Cell Mammals Maps Mass Spectrum Analysis Mediating Modification Molecular Monoubiquitination Mosaicism Mus Mutation Myosin ATPase Myosin Type II Nonmuscle Myosin Type IIA Organ PCDH15 gene Pathologic Phenotype Play Post-Translational Protein Processing Property Proteins Rapid screening Resources Retina Role Sensory Site Syndrome System Technology Testing Time Tissues Ubiquitination Usher Proteins Usher Syndrome Usher Syndrome Type 1 Variant Vertebrates Work X Chromosome base conditional mutant deaf deafness fly gene function hearing impairment human disease knock-down mechanotransduction mutant non-muscle myosin protein function rat Pres protein screening stable isotope ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Genetic screens in mice and the naturally occurring genetic variation in humans have provided a valuable resource to identify genes implicated in hair cell function and deafness. Two examples is the identification of genes involved in Usher syndrome, the most common form of deaf-blindness, and the role of Myh9 in both syndromic and non-syndromic deafness. We have used the power of Drosophila genetics to identify new genes involved in hearing and deafness. The auditory organs of Drosophila and vertebrates have a number of molecular and functional similarities despite being widely separated in evolutionary time. We identified mutations in Ubr3, an E3 ubiquitin ligase, that cause a physical detachment of the sensory components of Johnston's organ from the fly antenna. Strikingly, this phenotype is identical to that seen in mutations in Drosophila Myosin VIIa. Since Myosin VIIa mutations in humans cause Usher Syndrome type IB, it is possible that Ubr3 may regulate Myosin VIIa function in invertebrates and vertebrates. Our data suggest that Ubr3 genetically interacts with Myosin VIIa and Ubr3 and Myosin VIIa physically and genetically interact with Drosophila homologues of two other Usher syndrome proteins, PCDH15 and Sans. However, we have found that Ubr3 does not modify Myosin VIIa, but instead mono-ubiquitinates non-muscle Myosin II. This modification increase an interaction between the two myosins, and fine-tuning the level of this interaction appears critical for Myosin VIIa function. In the present proposal, we will expand on the use of Drosophila as a model system to understand deafness by characterizing the roles of Ubr3, Myosin II and Myosin VIIa in hearing in Drosophila (Aim 1). We will then test the function of Ubr3 in the development and function of mouse hair cells, and will test whether the interaction of Myosin II and Myosin VIIa is conserved in mice (Aim 2). Finally, we will carry out a genetic screen of 3000 Drosophila genes that may be involved in human disease using newly developed protein knockdown technology to identify genes that play a role in hearing in Drosophila (Aim 3).

项目摘要小鼠的遗传筛选和人类自然存在的遗传变异提供了宝贵的资源以识别与毛细胞功能和耳聋有关的基因。两个例子是识别参与Usher综合征的基因，聋盲的最常见形式，MyH9在两者中的作用综合征和非综合性耳聋。我们已经使用果蝇遗传学的力量来识别新的涉及听力和耳聋的基因。果蝇和脊椎动物的听觉器官有很多分子和功能相似性，尽管在进化时广泛分离。我们确定了 UBR3（E3泛素连接酶）中的突变，导致物理分离的感觉成分 Johnston的器官来自苍蝇天线。令人惊讶的是，这种表型与在突变中看到的表型相同果蝇肌球蛋白VIIA。由于人类中的肌球蛋白VIIA突变引起了usher综合征IB，因此有可能 UBR3可以调节无脊椎动物和脊椎动物中的肌球蛋白VIIA功能。我们的数据表明UBR3 遗传与肌球蛋白VIIA和UBR3以及肌球蛋白VIIA在物理和遗传上相互作用另外两种Usher综合征蛋白PCDH15和SANS的果蝇同源物。但是，我们发现该UBR3不会改变肌球蛋白VIIA，而是单次泛素化的非肌肉肌球蛋白II。此修改增加两种肌动物之间的相互作用，微调这种相互作用的水平似乎至关重要肌球蛋白VIIA功能。在本提案中，我们将扩展使用果蝇作为模型系统，以了解通过表征了UBR3，肌球蛋白II和肌球蛋白在果蝇中听力的作用（AIM 1）。然后我们将测试 UBR3在小鼠毛细胞的开发和功能中的功能，并将测试是否相互作用肌球蛋白II和肌球蛋白VIIA在小鼠中是保守的（AIM 2）。最后，我们将执行3000个遗传屏幕果蝇基因可能使用新开发的蛋白质敲低参与人类疾病鉴定在果蝇中听力中发挥作用的基因（AIM 3）。