CFTR/Regulation of CL Secretion in Normal and CF Airways

CFTR/正常气道和 CF 气道中 CL 分泌的调节

• 批准号: 7824134
• 负责人: William B. Guggino
• 金额: $ 0.82万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2009-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7824134
• 关键词: Abbreviations; Address; Affect; Area; Bacterial Toxins; Binding; Budgets; Cell membrane; Cells; Clostridium difficile; Critical Pathways; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Diarrhea; Disease; Environment; Epithelial; Epithelial Cells; GTPase-Activating Proteins; Goals; Golgi Apparatus; Golgi Targeting; Grant; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Human; Infection; Investigation; Lead; Ligands; Lung; Lysosomes; Macromolecular Complexes; Mediating; Membrane; Monomeric GTP-Binding Proteins; Pharmaceutical Preparations; Phosphoproteins; Physiological; Physiological Processes; Plasma; Play; Process; Protein Binding; Proteins; Pseudomonas Infections; Recycling; Regulation; Research Personnel; Role; Signal Transduction; Small Interfering RNA; Surface; System; Tight Junctions; Tissues; Toxin; World Health; base; cystic fibrosis airway; exoenzyme; ezrin; insight; member; moesin; new technology; novel; programs; protein function; radixin protein; rho; rho GTP-Binding Proteins; sodium-hydrogen exchanger regulatory factor; syntaxin 6; trafficking; trans-Golgi Network

DESCRIPTION (provided by applicant): The long-term goal of the grant over the past 15 years has been to understand both the Cl- channel and regulatory functions of CFTR. In the previous budget period, we focused specifically on the role of the PDZ binding domain and asked whether CFTR is a member of a macromolecular complex. We discovered a new protein, CAL that tethers CFTR within the Golgi and targets CFTR for degradation in the lysosome. We also studied NHE-RF and CAP70 how these PDZ domain-containing proteins function in relation to CAL, thus addressing the issue of why multiple PDZ domain-containing proteins bind to CFTR. The present proposal posits that the role of the interaction of CFTR with the PDZ domain of CAL is to regulate the amount of CFTR at the plasma either by tethering it at the Golgi, and targeting CFTR for degradation or allowing it to process to the plasma membrane. The proposal hypothesizes further, that the CAL-mediated decision whether CFTR is destined to move to the plasma membrane or not is determined by two associated proteins that bind to CAL, TC10 and syntaxin 6. The grant suggests further that the CAL-CFTR interaction with these two associated proteins has an important role in how CFTR responds to bacterial toxins that lead to diarrheal diseases and infection in the airways. The overall goal is to address the following three questions: Is the trafficking of mature CFTR to the plasma membrane regulated by TC10? Is the trafficking of mature CFTR to the plasma membrane regulated by Syntaxin6? How do bacterial toxins function in the trafficking of CFTR to the plasma membrane? Relevance: The proposal represents a new area of investigation with implications both for our understanding of diarrheal diseases involving toxins that affect Rho-GTPases and for Pseudomonas infection in lung where the toxin ExoS, may alter CFTR trafficking to the membrane. Once the critical pathways are identified and their roles established, it may be possible to develop drugs to therapeutically alter the switching mechanisms that determine CFTR's fate.

描述(由申请人提供)：在过去的15年里，该基金的长期目标一直是了解CFTR的氯离子通道和调节功能。在上一个预算期，我们特别关注PDZ结合域的作用，并询问CFTR是否是大分子复合体的成员。我们发现了一种新的蛋白质，CAL，它在高尔基体内捆绑CFTR，并在溶酶体中针对CFTR进行降解。我们还研究了NHE-RF和CAP70这些含有PDZ结构域的蛋白质如何与CAL相关的功能，从而解决了为什么多个含有PDZ结构域的蛋白质与CFTR结合的问题。目前的建议假设，CFTR与CAL的PDZ结构域的相互作用的作用是调节血浆中CFTR的量，要么将其拴在高尔基体上，要么靶向CFTR降解，要么允许它加工到质膜上。该提案进一步假设，CAL介导的决定CFTR是否注定要移动到质膜由两个与CAL结合的相关蛋白TC10和Synaxin 6决定。该研究进一步表明，CAL-CFTR与这两个相关蛋白的相互作用在CFTR对导致腹泻疾病和呼吸道感染的细菌毒素的反应中具有重要作用。总体目标是解决以下三个问题：成熟CFTR向质膜的运输是否受TC10的调控？成熟的cftr转运到质膜上是否受Synaxin6的调节？细菌毒素如何在CFTR向质膜转运过程中发挥作用？ 相关性：该提案代表了一个新的调查领域，对我们理解涉及影响Rho-GTP酶的毒素的腹泻疾病和肺部的假单胞菌感染都有影响，其中毒素EXOS可能改变CFTR向膜的运输。一旦确定了关键途径并确定了它们的作用，就有可能开发药物来从治疗上改变决定CFTR命运的转换机制。

项目成果

Effect of hypoxia on endothelin-1 production by pulmonary vascular endothelial cells.

缺氧对肺血管内皮细胞产生内皮素-1 的影响。

• DOI: 10.1016/0167-4889(92)90033-8
• 发表时间: 1992
• 期刊: Biochimica et biophysica acta
• 作者: Wiebke,JL;Montrose-Rafizadeh,C;Zeitlin,PL;Guggino,WB
• 通讯作者: Guggino,WB

Estrogen modulates ClC-2 chloride channel gene expression in rat kidney.

雌激素调节大鼠肾脏中 ClC-2 氯离子通道基因的表达。

• DOI: 10.1007/s00424-003-1095-y
• 发表时间: 2003
• 期刊: Pflugers Archiv : European journal of physiology
• 作者: Nascimento,DanielleS;Reis,CarlosU;Goldenberg,ReginaC;Ortiga-Carvalho,TâniaM;Pazos-Moura,CarmenC;Guggino,SandraE;Guggino,WilliamB;Morales,MarceloM
• 通讯作者: Morales,MarceloM

An improved system for packaging recombinant adeno-associated virus vectors capable of in vivo transduction.

• 发表时间: 1995
• 期刊: Gene therapy
• 影响因子: 5.1
• 作者: Terence R. Flotte;Ximena Barraza-Ortiz;R. Solow;S. Afione;B. Carter;W. Guggino

The GAP portion of Pseudomonas aeruginosa type III secreted toxin ExoS upregulates total and surface levels of wild type CFTR.

III 型铜绿假单胞菌分泌的毒素 ExoS 的 GAP 部分上调野生型 CFTR 的总水平和表面水平。

• DOI: 10.1159/000343357
• 发表时间: 2013
• 期刊: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
• 作者: Tukaye,DeepaliN;Kwon,Sang-Ho;Guggino,WiliamB
• 通讯作者: Guggino,WiliamB

Targeting CAL as a negative regulator of DeltaF508-CFTR cell-surface expression: an RNA interference and structure-based mutagenetic approach.

• DOI: 10.1074/jbc.m611049200
• 发表时间: 2007-03
• 期刊: The Journal of biological chemistry
• 作者: Michael Wolde;Abigail Fellows;Jie Cheng;Aleksandr Kivenson;B. Coutermarsh;L. Talebian;K. Karlson;A. Piserchio;D. Mierke;B. Stanton;W. Guggino;D. Madden