Biochemical Genetics of Carbonic Anhydrase Deficiencies

描述(申请人提供)：这项研究的主要目标是研究碳酸酐酶基因家族的功能基因组学，以确定单个成员对健康和疾病的重要性。已知的16种碳酸酐酶(CA)及其相关蛋白在呼吸、骨吸收、肾酸化、糖异生、信号转导、脑脊液和胃酸的形成等多种生理过程中发挥重要作用。我们有五个具体目标： 1)对新发现的碳酸酐酶XLV的性质和功能基因组学特性进行了研究。 2)鉴定CA IV基因敲除小鼠和CA IV/CA XIV双重基因敲除的表型后果。 3)验证CA IV信号序列突变是显性遗传性视网膜色素变性(RP17q)的潜在缺陷的假设。 4)鉴定线粒体碳酸氢酶CA-VA和CA-VB的性质和功能基因组学。 5)建立转基因小鼠模型，检测CA I/阴离子交换“代谢蛋白”在整个动物体内的功能重要性。 自从我们最初发现CA II缺乏综合症以来，我们寻求对一个具有强大生产力记录的项目的重新支持。这种疾病影响骨骼、大脑和肾脏，是第一个与CA缺乏有关的疾病。我们将使用各种生化、细胞生物学、免疫学和分子遗传学方法。通过定向突变，将产生单个CA缺陷的新的小鼠敲入和敲除小鼠模型。这些缺陷的功能后果将以多个生理测量为特征。在适当的情况下，单一缺乏CA的小鼠将通过杂交产生缺乏多个CA的小鼠。这些研究将加强我们对单个CA如何促进正常生理的理解，以及单个和双CA缺陷如何导致疾病。所寻求的答案具有基本意义，应该为CA缺乏症的新治疗方法提供信息。

DESCRIPTION (provided by applicant): The broad goal of this research is to study the functional genomics of the carbonic anhydrase gene family to determine the importance of individual members to health and disease. The sixteen known carbonic anhydrases (CAs) and CA-related proteins play important roles in diverse physiological processes including respiration, bone resorption, renal acidification, gluconeogenesis, signal transduction, and formation of cerebrospinal fluid and gastric acid. We have five specific aims: 1) Characterize the properties and functional genomics of newly discovered carbonic anhydrase XlV. 2) Characterize the CA IV knockout mouse and the phenotypic consequences of the CA IV/CA XIV double knockout. 3) Test the hypothesis that a signal sequence mutation in CA IV is the underlying defect in dominantly inherited retinitis pigmentosa (RP17q). 4) Characterize the properties and functional genomics of mitochondrial carbonic anhydrases CA VA and CA VB. 5) Create transgenic mouse models to test the functional importance of the CA I I/anion-exchanger "metabolon" in the whole animal. We seek renewed support for a program with a strong record of productivity since our initial discovery of the CA II deficiency syndrome. This disease affects bone, brain, and kidney and was the first disease associated with a CA deficiency. We will use a variety of biochemical, cell biological, immunological, and molecular genetic approaches. Novel mouse knock-in and knockout mouse models of individual CA deficiencies will be produced by targeted mutagenesis. Functional consequences of these deficiencies will be characterized by multiple physiological measurements. Mice deficient for multiple CAs will be produced, where appropriate, by intercrosses between singly deficient mice. These studies should enhance our understanding of how individual CAs contribute to normal physiology and how single and double CA deficiencies produce disease. The answers sought have fundamental significance, and should provide information leading to novel therapeutic approaches to CA deficiency.