KINESIN IN PHOTORECEPTOR CELLS

描述(由申请人提供)：本项目的长期总体目标 研究是为了帮助理解细胞机制在 光感受器细胞，尤其是那些参与细胞周转的细胞 光传导膜对光感受器细胞的活性很重要。这个 提议的计划是专注于了解激动素II在 感光细胞。在构成本提案基础的一项研究中， 我们研究了光感受器特异性基因敲除小鼠的光感受器细胞。 KIF3a基因，它是运动蛋白II的一个必需的运动亚基。 由一个杂三聚体组成，带有两个马达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基因敲除。 携带由IRBP启动子驱动的Cre。这项研究表明， 肌动蛋白II是成熟蛋白正常运输所必需的 光感受器细胞，是光感受器细胞生存所必需的。 在这项拟议的研究中，我们的目标是检验由这项研究产生的假设， 关于激动素II在蛋白质转运和光感受器中的作用 细胞退化。 我们将测试是否有任何编码激动素II亚单位的基因可以 对遗传性感光细胞变性负责。我们将探索 光感受器激动素II分子相互作用的测试 到目前为止，我们的基因敲除研究表明，蛋白质与蛋白质之间的相互作用。我们 将改进我们目前选择性敲除感光细胞的方法 KIF3a通过生成一种更有效的方式来交付Cre，因此理想情况下KIF3a 几乎所有的光感受器都可以同时去除。我们将使用这个 一种改进的方法来检验关于Kinesin II功能的不同假设 感光细胞。最后，我们将测试arrestin绑定的重要性 光感受器细胞死亡中的磷酸化视紫质。 这项拟议的研究与蛋白质如何传递的谜团有关 光感受器细胞的外节，到一般的功能 激动素，以及光感受器退化。光感受器变性，这是 是由各种不同基因的突变引起的(许多基因还没有发生 (已确认)，是人类失明的主要原因。

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.