KINESIN IN PHOTORECEPTOR CELLS

感光细胞中的驱动蛋白

• 批准号: 6318855
• 负责人: DAVID S WILLIAMS
• 金额: $ 27.83万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6318855
• 关键词: arrestins; gene expression; gene mutation; gene targeting; genetically modified animals; kinesin; laboratory mouse; protein binding; protein protein interaction; protein structure function; protein transport; retina degeneration; rhodopsin; uvea ciliary body; visual photoreceptor; visual phototransduction

DESCRIPTION (provided by applicant): The long-term overall goal of this research is to help in the understanding of cellular mechanisms in photoreceptor cells, especially those involved in turnover of the phototransductive membrane and important for photoreceptor cell viability. The proposed plan is to focus on understanding the role of kinesin II in photoreceptor cells. In a study that forms the basis for the present proposal, we studied photoreceptor cells in mice with photoreceptor-specific knock-out of the gene for KIF3a, an obligatory motor subunit of kinesin II. (Kinesin II consists of a heterotrimer, with two motor subunits and an accessory protein.) Selective knock-out was achieved using the cre-lox system, with transgenic mice carrying cre that was driven by the IRBP promoter. This study demonstrated that kinesin II is required for normal transport of proteins within mature photoreceptor cells and is essential for photoreceptor cell viability. In the proposed research, we aim to test hypotheses generated from this study, concerning the roles of kinesin II in protein transport and in photoreceptor cell degeneration. We will test whether any of the genes encoding the subunits of kinesin II could be responsible for inherited photoreceptor cell degeneration. We will explore the molecular interactions of photoreceptor kinesin II by testing protein-protein interactions suggested from our knock-out studies thus far. We will improve upon our current method of selective knock-out of photoreceptor KIF3a by generating a more effective way to deliver cre, so that ideally KIF3a can be removed in nearly all photoreceptors at the same time. We will use this improved method to test different hypotheses about kinesin II function in photoreceptor cells. Finally, we will test the importance of arrestin-bound phosphorylated rhodopsin in photoreceptor cell death. The proposed research is pertinent to the mystery of how proteins are delivered to the outer segment of photoreceptor cells, to the general function of kinesins, and to photoreceptor degeneration. Photoreceptor degeneration, which is caused by mutations in a variety of different genes (many yet to be identified), is a major cause of human blindness.

描述（由申请人提供）：本项目的长期总体目标 研究旨在帮助理解细胞机制 感光细胞，尤其是那些参与感光细胞更新的细胞 光导膜对感光细胞的活力很重要。这 拟议的计划是重点了解驱动蛋白 II 在 感光细胞。在构成本提案基础的一项研究中， 我们研究了小鼠感光细胞的感光细胞特异性敲除 KIF3a 的基因，KIF3a 是驱动蛋白 II 的必需运动亚基。 （驱动蛋白 II 由异源三聚体组成，具有两个运动亚基和一个辅助蛋白。） 使用 cre-lox 系统对转基因小鼠进行选择性敲除 携带由 IRBP 启动子驱动的 cre。这项研究表明 驱动蛋白 II 是成熟体内蛋白质正常运输所必需的 感光细胞，对于感光细胞的活力至关重要。 在拟议的研究中，我们的目标是测试本研究中产生的假设， 关于驱动蛋白 II 在蛋白质转运和光感受器中的作用 细胞变性。 我们将测试编码驱动蛋白 II 亚基的任何基因是否可以 负责遗传性感光细胞退化。我们将探索 通过测试光感受器驱动蛋白 II 的分子相互作用 迄今为止我们的敲除研究表明蛋白质-蛋白质相互作用。我们 将改进我们目前选择性敲除光感受器的方法 KIF3a 通过生成更有效的方式来传递 cre，以便理想情况下 KIF3a 几乎可以同时去除所有光感受器中的光感受器。我们将使用这个 测试有关驱动蛋白 II 功能的不同假设的改进方法 感光细胞。最后，我们将测试抑制蛋白结合的重要性 磷酸化视紫质在感光细胞死亡中的作用。 拟议的研究与蛋白质如何传递的奥秘有关 感光细胞的外段，一般功能 驱动蛋白和光感受器变性。光感受器变性， 是由多种不同基因的突变引起的（许多基因尚未被证实） 已确定），是人类失明的主要原因。