Molecular bases of BK channel function and localization

BK通道功能和定位的分子基础

基本信息

批准号：
8197884
负责人：
ZHAO-WEN WANG
金额：
$ 30.49万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-12-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8197884
关键词：
Biological Assay Biotin Caenorhabditis elegans Cerebellar Ataxia Chimeric Proteins Chromosome Mapping Disease Disease Management Drosophila genus Engineering Epilepsy Erectile dysfunction Excretory function Fluorescence Functional disorder Future Genes Genetic Screening Genome Goals Haploidy Health Homologous Gene Human Hypertension Immunohistochemistry Interneurons Kidney Knowledge Label Mammals Membrane Mink Molecular Molecular Genetics Muscle Muscle Cells Mutation Names Nervous system structure Neuromuscular Junction Neurons Overactive Bladder Paroxysmal Dyskinesias Peptides Phenotype Physiological Presynaptic Terminals Property Protein Isoforms Proteins Research Screening procedure Site Surface Synapses System Testing Western Blotting Xenopus oocyte base density gain of function genetic regulatory protein glomerular filtration hearing impairment in vivo large-conductance calcium-activated potassium channels loss of function mutation mutant neurotransmitter release novel postsynaptic presynaptic programs research study voltage

项目摘要

DESCRIPTION (provided by applicant): BK channels are almost ubiquitously expressed and perform many important physiological functions. Dysfunction of the channel causes a variety of diseases, including epilepsy, progressive hearing loss, cerebellar ataxia, and hypertension. The function of BK channels depends on proper subcellular localization and interactions with auxiliary or regulatory proteins. However, the molecular basis of BK channel subcellular localization is unknown and only limited knowledge exists about BK channel auxiliary/regulatory proteins. The powerful molecular genetics of C. elegans is explored to identify proteins important to BK channel function or subcellular localization. In preliminary studies, mutants of three genes (named as bkip-1, bkip-2 and bkip-3) were isolated as suppressors of a lethargic phenotype caused by expressing a gain-of-function (gf) isoform of SLO-1, the C. elegans BK channel. Phenotypes of these mutants included increased neurotransmitter release (bkip-1) and SLO-1 mislocalization (bkip-2 and bkip-3). bkip-1 and bkip-2 were found to encode novel BK channel- interacting proteins whereas bkip-3 remains to be identified. BKIP-1 showed several effects on SLO-1 functional properties when analyzed in a heterologous expression system. This proposal is to test the hypotheses that the three BKIPs are important to SLO-1 function and/or subcellular localization in vivo and that there are other functionally related proteins in C. elegans. The specific aims of this proposal are: (1) determine how BKIP-1 regulates SLO-1 function; (2) determine how BKIP-2 and BKIP-3 control SLO-1 subcellular localization and/or function, and (3) isolate and identify additional mutants that suppress the lethargic phenotype caused by SLO-1(gf), which will be analyzed in future studies. The long-term goal is to elucidate the molecular basis of BK channel function and subcellular localization. PUBLIC HEALTH RELEVANCE: Mutations of the BK channel cause a variety of diseases, including epilepsy, hypertension, progressive hearing loss, cerebellar ataxia, overactive bladder, penile erectile dysfunction, impaired renal glomerular filtration and K+ excretion, and paroxysmal dyskinesia. The proposed research program may identify novel proteins that are important to BK channel function and/or subcellular localization in vivo. Such information is potentially of great value to understanding the molecular bases of BK channel-related diseases, and to identifying candidate pharmacological targets for the treatment and management of these diseases.

描述（由申请人提供）：BK通道几乎普遍表达并执行许多重要的生理功能。通道的功能障碍会引起各种疾病，包括癫痫，进行性听力损失，小脑共济失调和高血压。 BK通道的功能取决于适当的亚细胞定位以及与辅助或调节蛋白的相互作用。但是，BK通道亚细胞定位的分子基础尚不清楚，并且仅存在有关BK通道辅助/调节蛋白的知识有限。探索了秀丽隐杆线虫的强大分子遗传学，以鉴定对BK通道功能或亚细胞定位重要的蛋白质。在初步研究中，将三个基因的突变体（称为BKIP-1，BKIP-2和BKIP-3）分离为作为表达功能障碍（GF）Slo-1的嗜and c. equans BK通道引起的嗜睡表型的抑制。这些突变体的表型包括增加的神经递质释放（BKIP-1）和SLO-1错误定位（BKIP-2和BKIP-3）。发现BKIP-1和BKIP-2编码新型BK通道相互作用蛋白，而BKIP-3仍有待鉴定。当在异源表达系统中分析时，BKIP-1对SLO-1功能特性显示出几种影响。该建议是测试三个BKIP对体内SLO-1功能和/或亚细胞定位至关重要的假设，并且在秀丽隐杆线虫中还有其他功能相关的蛋白质。该提案的具体目的是：（1）确定BKIP-1如何调节SLO-1功能；（2）确定BKIP-2和BKIP-3控制SLO-1亚细胞定位和/或功能如何，以及（3）隔离并鉴定抑制由SLO-1（GF）引起的嗜睡表型的其他突变体，这将在未来的研究中进行分析。长期目标是阐明BK通道功能和亚细胞定位的分子基础。公共卫生相关性：BK通道的突变会引起多种疾病，包括癫痫，高血压，进行性听力损失，小脑共济失调，过度活跃的膀胱，阴茎勃起功能障碍，肾肾小球过滤受损，K+++++ ESKCRETION和PAROXYSMAL DYSKINESIA。提出的研究计划可以鉴定出对BK通道功能和/或体内亚细胞定位很重要的新型蛋白质。这些信息对于理解与BK通道相关疾病的分子基础以及确定候选药理学靶标的治疗和治疗这些疾病具有巨大价值。