The Role of Ribosome Biogenesis Factors in Liver Cirrhosis

核糖体生物发生因子在肝硬化中的作用

• 批准号: 8266016
• 负责人: EMILY Frances FREED
• 金额: $ 0.6万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8266016
• 关键词: Affect; Affinity Chromatography; Alcohol-Induced Disorders; Alcoholic Liver Diseases; Antibodies; Area; Biogenesis; Biological Assay; Biopsy Specimen; Cell Nucleolus; Cell division; Cell physiology; Cessation of life; Childhood; Cirrhosis; Co-Immunoprecipitations; Defect; Development; Disease; Eukaryota; Fluorescence Microscopy; Goals; Growth; Growth and Development function; Health; Hepatocyte; Homologous Gene; Human; Immunofluorescence Microscopy; Lead; Learning; Liver; Liver Cirrhosis; Liver diseases; Malignant neoplasm of liver; Methods; Missense Mutation; Mutation; North American Indians; Northern Blotting; Pathology; Primary carcinoma of the liver cells; Protein Biosynthesis; Proteins; Reporting; Research; Ribosomal RNA; Ribosomes; Risk; Risk Factors; Role; Testing; Tissue Sample; Tissues; United States; Yeasts; cDNA Library; cancer cell; cell growth; cell injury; human disease; insight; knock-down; liver biopsy; liver cell proliferation; novel; prevent; research study; response; yeast two hybrid system

DESCRIPTION (provided by applicant): Ribosomes, the cellular factories responsible for making proteins, are essential for cell growth and division. Although a complete loss of ribosome biogenesis is expected to be lethal, dysfunctional ribosome biogenesis leads to a variety of human diseases. Recently, a missense mutation in the ribosome biogenesis protein, Cirhin, was reported to cause North American Indian childhood cirrhosis (NAIC). This suggests that ribosome biogenesis may be involved in other forms of cirrhosis, including alcoholic liver disease (ALD). The overall goal of this proposal is to examine the relationship between ribosome biogenesis and cirrhosis. The first aim of this proposal is to identify novel proteins that are involved in ribosome biogenesis in liver cells by determining the protein interaction partners of Cirhin through both a yeast-two hybrid screen and affinity purification followed by mass spectrometery. One candidate protein, NOL11 has already been identified. The second aim of this proposal is therefore to characterize any novel Cirhin-interacting proteins, including NOL11, and analyze their role in ribosome biogenesis. Finally, the third aim is to begin to elucidate the role of ribosome biogenesis in ALD. This will be accomplished by determining expression levels of ribosome biogenesis factors, including Cirhin, NOL11, and any other newly identified proteins, using immunofluorescence microscopy of liver tissue samples encompassing the whole spectrum of alcohol-induced liver injury. Since these proteins are required for ribosome biogenesis, their expression levels provide an easy indicator of whether ribosome biogenesis is occurring. Through these experiments, we will gain insight into how ribosome biogenesis affects both normal liver growth and the development of liver pathology.

描述（由申请人提供）：核糖体是负责制造蛋白质的细胞工厂，对于细胞生长和分裂至关重要。尽管核糖体生物发生的完全丧失预计会致命，但核糖体生物发生功能失调会导致多种人类疾病。最近，据报道核糖体生物发生蛋白 Cirhin 的错义突变导致北美印第安人儿童期肝硬化 (NAIC)。这表明核糖体生物发生可能与其他形式的肝硬化有关，包括酒精性肝病（ALD）。该提案的总体目标是检查核糖体生物发生与肝硬化之间的关系。该提案的第一个目的是通过酵母-两种杂交筛选和亲和纯化以及质谱分析来确定 Cirhin 的蛋白质相互作用伙伴，从而鉴定参与肝细胞核糖体生物合成的新型蛋白质。一种候选蛋白 NOL11 已经被鉴定出来。因此，该提案的第二个目标是表征任何新型 Cirhin 相互作用蛋白（包括 NOL11），并分析它们在核糖体生物发生中的作用。最后，第三个目标是开始阐明核糖体生物发生在 ALD 中的作用。这将通过使用涵盖酒精性肝损伤全谱的肝组织样本的免疫荧光显微镜确定核糖体生物合成因子（包括 Cirhin、NOL11 和任何其他新发现的蛋白质）的表达水平来实现。由于这些蛋白质是核糖体生物发生所必需的，因此它们的表达水平提供了核糖体生物发生是否发生的简单指示。通过这些实验，我们将深入了解核糖体生物发生如何影响正常肝脏生长和肝脏病理学的发展。