REGULATION OF GAMMA CARBOXYLASE GENE TRANSCRIPTION IN LIVER

肝脏中γ羧化酶基因转录的调控

• 批准号: 6657099
• 负责人: David A Roth
• 金额: $ 18.67万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6657099
• 关键词: age difference; aminoacid biosynthesis; clotting factor; developmental genetics; embryo /fetus cell /tissue; gamma carboxyglutamate; genetic mapping; genetic transcription; genetically modified animals; laboratory mouse; liver metabolism; mammalian embryology; nucleic acid sequence; transcription factor; vitamin K

The expression of the vitamin K-dependent carboxylase gene in liver is essential for Gla formation in the vitamin K-dependent proteins, and is therefore essential for hemostasis. The embryonic expression of the vitamin K-dependent carboxylase gene, in hepatic and extrahepatic tissues, is also essential for survival and normal development of the fetus. The tissue-specific expression of the carboxylase gene is developmentally regulated, yet the mechanism underlying its temperospatial regulation is unknown. DNA sequences in the 5' flanking region of the carboxylase gene that are important in the transcriptional control of the carboxylase have been identified by transient gene expression assays in cell line models of hepatocytes differentiation. The functions of these regions during hepatocyte development in vivo and their importance in the control of extrahepatic carboxylase gene expression is unknown. The objectives of this application are to elucidate the regulatory mechanisms that control the developmental expression of this essential gene in liver. Studies in this application are divided into two specific aims. In the first aim, transcription factors responsible for carboxylase gene expression will be isolated and analyzed. In the second aim, DNA sequences important in the developmental regulation of vitamin K-dependent carboxylase gene transcription in vivo will be identified using transgenic animal models. Regions of the carboxylase gene important in its in vivo expression will be identified by Dnase I hypersensitivity analysis of the carboxylase gene locus in its in vivo expression will be identified by Dnase I hypersensitivity analysis of the carboxylase gene locus in nuclei from immature and mature liver. Studies of DNA-protein interactions at regulation of Gla synthesis in developing and differentiated tissues. The findings of the proposed studies may provide important insights into pathogenesis of the warfarin embryopathy. Knowledge of the mechanisms that regulate hepatic and extrahepatic Gla-synthesis may also lead to alternative strategies for treatment and prevention of several diseases. Selective tissue-specific modulation of vitamin K-dependent protein synthesis may lead to novel anti-coagulation strategies important in the clinical management of thrombotic disorders.

维生素K依赖性羧化酶基因在肝脏中的表达对于维生素K依赖性蛋白的GLA形成至关重要，因此对于止血至关重要。肝和肝外组织中维生素K依赖性羧化酶基因的胚胎表达对于胎儿的存活和正常发育也是必不可少的。羧化酶基因的组织特异性表达在发育中受到调节，但是其温度空间调节的机制尚不清楚。羧化酶基因的5'侧翼区域中对羧化酶转录对照很重要的DNA序列已通过瞬时基因表达测定在肝细胞分化的细胞系模型中鉴定出来。这些区域在体内发育过程中的功能及其在控制肝外羧化酶基因表达中的重要性尚不清楚。该应用程序的目标是阐明控制肝脏中基因的发育表达的调节机制。该应用中的研究分为两个具体目标。在第一个目标中，将分离和分析负责羧化酶基因表达的转录因子。 在第二个目的中，使用转基因动物模型鉴定出体内维生素K依赖性羧化酶基因转录的发育调节重要的DNA序列。羧化酶基因在其体内表达中很重要的区域将通过DNase I的超敏反应分析在其体内表达中对羧化酶基因座的超敏反应分析，将通过DNase I I超敏反应分析对核中的羧化酶基因基因座的超敏性分析，来自核中的核中，来自核中的核和成熟肝脏中。 DNA-蛋白质相互作用在调节GLA合成和分化组织中的研究。拟议研究的发现可能会为华法林胚胎病的发病机理提供重要的见解。了解调节肝和肝外GLA合成的机制也可能导致治疗和预防多种疾病的替代策略。维生素K依赖性蛋白质合成的选择性组织特异性调节可能导致对血栓性疾病临床管理重要的新型抗凝策略。