REGULATION OF CILIATED CELL DIFFERENTIATION

纤毛细胞分化的调节

• 批准号: 6162176
• 负责人: L OSTROWSKI
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6162176
• 关键词: animal genetic material tag; cell differentiation; cell growth regulation; cilium; complementary DNA; dynein ATPase; enzyme activity; gene induction /repression; laboratory rat; molecular cloning; nucleic acid sequence; open reading frames; respiratory epithelium

Summary of Work: Ciliated cells line the surface of the airways, and by the coordinated beating of their cilia provide the force necessary for mucociliary clearance. Patients with primary ciliary dyskinesia (PCD) suffer from repeated respiratory infections. Ciliated cells are easily damaged by a wide range of air pollutants and pathogens. Once damaged, it is essential that ciliated cells be regenerated and mucociliary clearance restored. The main goal of our research is to understand the regulation of ciliogenesis. We have begun several projects to investigate the mechanisms regulating the expression of ciliated cell-specific genes. We have identified and cloned partial cDNAs for seven unique axonemal dyneins from rat tracheal epithelial cells (RTE). The expression of these dyneins correlates with the development of ciliated cells. Because these dyneins are very large, we are using a PCR based strategy to clone larger pieces of these genes in the 5' direction. We have successfully "walked" 4.5 kb (out of the expected 6-7 kb) on one of the more abundant dyneins, and have made progress on several others. In addition, we have identified two novel genes, KPL1 and KPL2, which are upregulated during ciliogenesis using differential display. The complete cDNA for KPL1 has been cloned and sequenced and codes for a predicted protein of 24 kD. This unique protein contains a pleckstrin homology domain, which indicates that KPL1 probably plays a role in a signal transduction pathway. We have sequenced 3.7 kb of the 7 kb KPL2 message, and so far identified an ATP binding site in the predicted open reading frame. This message is only expressed in tissues that contain ciliated cells.

工作总结：纤毛细胞排列在气道表面， 纤毛的协调跳动提供了必要的力量 粘膜纤毛清除率原发性纤毛运动障碍（PCD）患者 反复呼吸道感染。 纤毛细胞很容易 受到各种空气污染物和病原体的破坏。一旦损坏， 是必不可少的纤毛细胞再生和粘膜纤毛清除 恢复. 我们研究的主要目的是了解 纤毛发生我们已经开始了几个项目来调查 调节纤毛细胞特异性基因表达的机制。我们 已经鉴定并克隆了七种独特的轴丝的部分cDNA 动力蛋白来自大鼠气管上皮细胞（RTE）。表达这些 动力蛋白与纤毛细胞的发育相关。因为这些 动力蛋白非常大，我们正在使用基于PCR的策略来克隆更大的 这些基因的片段在5'方向。我们已经成功地“走”了 4.5 kb（超出预期的6-7 kb）在一个更丰富的动力蛋白上， 并在其他几个方面取得了进展。此外，我们还发现， 两个新的基因，KPL 1和KPL 2，在纤毛发生过程中上调 使用差分显示。克隆了KPL 1的完整cDNA 测序并编码24 kD的预测蛋白。这种独特 蛋白含有普列克底物蛋白同源结构域，这表明KPL 1 可能在信号转导通路中起作用。我们已经测序了 3.7 7 kb KPL 2信息中的1 kb，到目前为止确定了ATP结合 在预测的开放阅读框架中的位点。这条信息只表达了 在含有纤毛细胞的组织中。