Developmental Program and Genetic Disease by Six family genes.

六个家族基因的发育计划和遗传疾病。

基本信息

批准号：
12470029
负责人：
KAWAKAMI Kiyoshi
金额：
$ 9.09万
依托单位：
Jichi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

This study aims to reveal roles of Six family genes in development, to elucidate gene network including Six and involvement of Six genes in pathology of myotonic dystrophy (DM1). We performed analyses of Six gene defective mice, screening of target genes of Six proteins and analyses of molecular function of Eya and Dach protein that are cooperative factors of Six protein.1 Six4/Six5 double knockout mice die within several hours after birth but we could not find any apparent anatomical anomalies. Six1 gene defective mice die just after birth and showed defective formation of inner ear, nose, kidney and thymus. The morphological abnormalities were noted from E10-11. Six1 gene is suggested to be essential for the formation of these organs.2 Target genes of Six5 proteins were identified. Transcription factors, signaling molecules and its receptors that are expressed during mesoderm differentiation were identified as putative target in P19 cells. Transcription factors and signaling molecules in nervous systems, transporters and receptor proteins of neural transmitters. In myoblasts, genes including myogenin, myosin, troponin, acetylcholine receptors that arespecific to skeletal muscle were identified. In lens epithelial cells, genes that had been shown to be involved in cataractogenesis were identified. It is suggested that altered regulation of these target genes leads to some symptoms of DM1.3 We revealed that cooperative activation by GAL4-Eya and Dach is mediated through CBP. CBP bound to an immobilized chromatin template only in the presence of both GAL4-Eya and Dach. We also found that Dach can bind to chromatin as well as DNA regardless of the presence of GAL4-Eya protein. The binding affinity to chromatin was higher than that to naked DNA. The conserved DD1 domain of Dach is responsible for the DNA binding activity.

这项研究旨在揭示六个家庭基因在发育中的作用，以阐明基因网络，包括六个基因和六个基因参与肌发育症病理学（DM1）。我们对六只基因有缺陷的小鼠进行了分析，对六种蛋白质的靶基因进行筛选，并分析Eya和Dach蛋白的分子功能，这是六种蛋白的合作因子。1六六/六个双重敲除小鼠在出生后几个小时内死亡，但我们可以找不到任何明显的解剖异常。 Six1基因有缺陷的小鼠出生后死亡，并显示出内耳，鼻子，肾脏和胸腺的缺陷。从E10-11注意到形态异常。认为SIX1基因对这些器官的形成至关重要。2鉴定出六个蛋白质的靶基因。在中胚层分化过程中表达的转录因子，信号分子及其受体被确定为p19细胞中的推定靶标。神经发射器的神经系统，转运蛋白和受体蛋白中的转录因子和信号分子。在肌细胞中，鉴定了肌蛋白，肌球蛋白，肌钙蛋白，乙酰胆碱受体，包括肌蛋白，乙酰胆碱受体。在晶状体上皮细胞中，已经鉴定出已证明与白内迫切发生有关的基因。有人提出，这些靶基因的调节改变导致DM1.3的某些症状，我们揭示了GAL4-EYA和DACH的合作激活是通过CBP介导的。 CBP仅在GAL4-EYA和DACH存在下与固定的染色质模板结合。我们还发现，DACH可以与染色质以及DNA结合，而与GAL4-EYA蛋白的存在无关。与染色质的结合亲和力高于裸体DNA。 DACH的保守DD1结构域负责DNA结合活性。