Regulation of Signaling of PKD2 Cation Channel in Sperm

精子中 PKD2 阳离子通道信号传导的调节

基本信息

批准号：
7190558
负责人：
XIANGYI LU
金额：
$ 29.96万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7190558
关键词：
Accounting Adaptor Signaling Protein Affect Aneurysm Antibodies Autosomal Dominant Polycystic Kidney Biochemical Pathway Biological Assay Biological Process Blood Vessels C-terminal Calcium Candidate Disease Gene Cations Cell Volumes Cell membrane Cells Chemicals Cilia Coiled-Coil Domain Complex Culture Media Cultured Cells Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cyst Cystic kidney Decompression Sickness Defect Disease Drosophila genus Elements End stage renal failure Endoplasmic Reticulum Epithelial Event Family Female Frequencies GTP-Binding Proteins Gel Genes Genetic Goals Grant Growth Hereditary Disease Homeostasis Hypertension In Vitro Integral Membrane Protein Intercellular Junctions Ionophores Kidney Knock-out Lead Left Localized Mammals Maps Mechanics Mediating Methods Modeling Molecular Movement Mus Mutate Mutation Nodal Numbers PKD2 protein Patch-Clamp Techniques Pathology Pathway interactions Patients Phenocopy Phenotype Phosphoproteins Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Play Polycystic Kidney Diseases Post-Translational Protein Processing Property Protein Kinase Proteins Proteomics Regulation Renal tubule structure Research Research Personnel Role Sampling Signal Pathway Signal Transduction Site Sperm Motility Stimulus Structure System Tail Tertiary Protein Structure Testing Therapeutic Intervention Work cell motility cell type extracellular fluid flow genetic analysis in vivo insight loss of function mortality mutant neuronal cell body novel prevent renal epithelium reproductive response sperm cell sperm function sperm protein tandem mass spectrometry trafficking

项目摘要

DESCRIPTION (provided by applicant): Autosomal dominant polycystic kidney disease (ADPKD) is characterized by kidney cyst formation that progresses to end-stage renal failure at a high frequency. The disease also encompasses systemic vascular complications such as hypertension and vascular aneurysms that increase the overall mortality of the disease. Mutations in two genes Pkd1 and Pkd2 account for most cases of ADPKD. Recent studies show that PKD2 functions as a calcium-activated, calcium-permeable cation channel; and PKD1 plays a role in regulating the PKD2 channel activity. A high concentration of PKD1 and PKD2 co-localizes on primary cilia of renal epithelia. The current working model is that cilium-associated PKD1/PKD2 protein complex mediates Ca2+ influx through cilia bending in response to fluid flow in the renal tubules. Loss of either PKD1 or PK02 leads to a common defect in ciliary Ca2+ influx and this is one of the contributing factors of cyst formation in the kidney. This proposal utilizes a novel ciliary model of PKD2 in Drosophila sperm to substantiate the current model of PKD2 as cilium-associated, mechano-/chemo-sensitive cation channel. We show that Drosophila PKD2 (CG6504) is a conserved cation channel of the PKD2 family that localizes to sperm flagellar cilia where it is required for responding to signals that regulate asymmetric flagellar motility and directional sperm movement. This Drosophila PKD2 function appears to be analogous to mammalian PKD2 function in the motile nodal cilia. Directional beating of nodal cilia, which contain PKD2, is required for left-right axis determination in mammals. The goals of this proposal are to elucidate PKD2 channel regulation and signaling through structure-function analyses of the Drosophila PKD2 protein, phosphoproteomic analyses of PKD2-dependent downstream protein phosphorylation. and genetic analyses of candidate kinases and phosphoproteins in the pathway. The proposed research will identify structural elements involved in PKD2 channel regulation and function in vivo in a ciliary context, and will also identify PKD2-dependent downstream phosphoproteins and use them to delineate a genetic pathway for PKD2 signaling and function.

描述（由申请人提供）：常染色体显性多囊性肾脏疾病（ADPKD）的特征是肾脏囊肿形成，该肾脏形成在高频下发展为终末期肾衰竭。该疾病还包括全身血管并发症，例如高血压和血管动脉瘤，增加了疾病的总体死亡率。在大多数ADPKD病例中，两个基因中的突变是PKD1和PKD2。最近的研究表明，PKD2充当钙激活的钙渗透阳离子通道。 PKD1在调节PKD2通道活性中起作用。高浓度的PKD1和PKD2在肾上皮的原发性纤毛上共定位。当前的工作模型是，与纤毛相关的PKD1/PKD2蛋白复合物介导了通过纤毛弯曲的Ca2+涌入，以响应肾小管中的流体流动。 PKD1或PK02的丧失导致睫状Ca2+流入的常见缺陷，这是肾脏囊肿形成的因素之一。该建议利用果蝇精子中PKD2的新型睫状模型来证实PKD2的当前模型作为纤毛相关的机械/化学敏感阳离子通道。我们表明，果蝇PKD2（CG6504）是PKD2家族的保守阳离子通道，它本地定位于精子鞭毛纤毛，在这里需要响应调节不对称的鞭毛运动和方向精子运动的信号。该果蝇PKD2功能似乎类似于在纤毛纤毛中的哺乳动物PKD2功能。哺乳动物中左右轴确定所必需的含有PKD2的节点纤毛的定向跳动。该提案的目标是通过果蝇PKD2蛋白的结构 - 功能分析，PKD2依赖性下游蛋白磷酸化的磷酸蛋白质组学分析来阐明PKD2通道调节和信号传导。途径中候选激酶和磷蛋白的遗传分析。拟议的研究将在纤毛环境中识别PKD2通道调节和功能中涉及的结构元素，还将识别依赖PKD2依赖性的下游磷蛋白，并使用它们来描述PKD2信号传导和功能的遗传途径。