Myosin XVA and Other Genes Essential for Stereocilia Morphogenesis

肌球蛋白 XVA 和其他对于立体纤毛形态发生至关重要的基因

基本信息

批准号：
9147428
负责人：
Thomas Friedman
金额：
$ 95.44万
依托单位：
NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9147428
关键词：
17p 17p11.2 Actins Alleles Alternative Splicing Apical Auditory system Binding Binding Proteins Biological Biological Assay Birds Bundling Cells Chromosomes Collaborations Complex Data Doctor of Philosophy EPS8 gene Endocrine Engineering Exons Family Family member Genes Goals Hair Cells Head Hearing Hearing Impaired Persons Heat-Shock Proteins 90 Human Inherited Insecta Kinetics Knockout Mice Laboratories Labyrinth Length Libraries Light Link Molecular Chaperones Molecular Motors Morphogenesis Motor Mus Mutate Mutation Myosin ATPase N-terminal National Heart, Lung, and Blood Institute National Institute on Deafness and Other Communication Disorders Neck Phenotype Play Population Protein Isoforms Proteins Publishing Reporting Research Design Role Stereocilium Supporting Cell Surface System Tail Tissues WHRN gene Yeasts base biophysical analysis biophysical properties cell type congenital deafness deafness hearing impairment mouse model mutant myosin XVA protein function yeast two hybrid system

项目摘要

Mutant alleles of mammalian myosin XVA cause profound congenital deafness DFNB3. In humans worldwide, mutations of MYO15A apepars to be a common mutated gene associated with nonsyndromic deafness, perhaps the 4th most common gene in which mutations are associated with congenital hearing loss. Myosins are actin-activated molecular motors that have a conserved head (motor) and neck (light chain binding motifs) and highly divergent tail domains. The MYO15A tail contains several motifs that are candidates for protein interaction motifs. To date, isoform 1 of myosin XVa is the largest of all reported vertebrate myosins. The N-terminus of myosin XVa is composed of 1,220 residues. We previously demonstrated that isoform 1 is necessary for hearing (Nal et al., 2007). As a collaboration with Drs. Sally Camper and Gregory Frolenkov, a mouse model has been developed that has a defective amino terminus which recapitulates the human phenotype (unpublished data). The identification of proteins that functionally interact with MYO15 may provide a means of determining the role of MYO15A in the auditory system. In addition, interacting proteins are themselves likely to play crucial roles in hearing and would be strong candidates for proteins encoded by other deafness loci. We are therefore using a yeast two hybrid system and MS screens to identify proteins that interact with the myosin XVA. Genes that encode poteins that interact with myosin XVA from these two screens will be further examined for biological relevance. Whirlin and EPS8 are reported partners of the tail domain of myosin XVa and are necessary for stereocilia elongation and staircase formation (Belyantseva et al. 2005). As a collaboration with Dr. Sellers (NHLBI), we are characterizing the biophysical properties of myosin XVa (Bird et al., 2014).

哺乳动物肌球蛋白 XVA 的突变等位基因导致严重的先天性耳聋 DFNB3。在全世界人类中，MYO15A 突变似乎是与非综合征性耳聋相关的常见突变基因，可能是与先天性听力损失相关的第四个最常见基因。肌球蛋白是肌动蛋白激活的分子马达，具有保守的头部（马达）和颈部（轻链结合基序）以及高度分化的尾部结构域。 MYO15A 尾部包含多个作为蛋白质相互作用基序候选的基序。迄今为止，肌球蛋白 XVa 的异构体 1 是所有已报道的脊椎动物肌球蛋白中最大的。肌球蛋白 XVa 的 N 末端由 1,220 个残基组成。我们之前证明异构体 1 对于听力是必需的 (Nal et al., 2007)。作为与博士的合作。 Sally Camper 和 Gregory Frolenkov 开发了一种小鼠模型，其氨基末端有缺陷，可以概括人类表型（未发表的数据）。与 MYO15 功能性相互作用的蛋白质的鉴定可能提供一种确定 MYO15A 在听觉系统中的作用的方法。此外，相互作用的蛋白质本身可能在听力中发挥关键作用，并且可能是其他耳聋基因座编码的蛋白质的有力候选者。因此，我们使用酵母两种杂交系统和 MS 筛选来鉴定与肌球蛋白 XVA 相互作用的蛋白质。将进一步检查这两个筛选中编码与肌球蛋白 XVA 相互作用的蛋白的基因的生物学相关性。据报道，Whirlin 和 EPS8 是肌球蛋白 XVa 尾部结构域的伙伴，对于静纤毛伸长和阶梯形成是必需的 (Belyantseva et al. 2005)。作为与 Sellers 博士 (NHLBI) 的合作，我们正在表征肌球蛋白 XVa 的生物物理特性 (Bird et al., 2014)。