International Research Fellowship Program: Determination of the Function of RoXaN I in Cellular and Rotavirus Biology

[401866]贝克尔国际研究奖学金计划使美国科学家和工程师能够到国外进行3至24个月的研究。该计划的奖励为联合研究提供了机会，并利用独特或互补的设施、专业知识和国外的实验条件。该奖项将支持Michelle M. Becker博士与Didier Poncet博士在法国伊维特河畔吉夫国家科学研究中心开展为期12个月的研究。RoXaN I是一种没有已知功能的细胞蛋白，通过与轮状病毒非结构蛋白NSP3的相互作用被发现。RoXaN存在于细胞核和细胞质中，并可能在这两个细胞区室之间移动，可能运输mRNA或调节翻译。这些实验旨在验证在轮状病毒感染期间RoXaN的正常功能被破坏以增强病毒复制的假设，并将提供有关轮状病毒复制机制和病毒控制细胞过程的新信息。由于RoXaN存在于细胞质和细胞核中，但不包含经典的核定位序列或核输出序列，因此使用共聚焦免疫荧光显微镜确定RoXaN在未感染和轮状病毒感染细胞中的亚细胞定位。RoXaN的缺失突变体被用来定义适当的亚细胞定位所必需的蛋白质结构域。这些实验的结果将确定RoXaN在细胞内的定位，以及蛋白质内对将RoXaN引导到正确的细胞室至关重要的区域。在RoXaN表达和不表达的情况下，将检测RoXaN对轮状病毒过程的影响。使用RNA干扰技术删除RoXaN mRNA的细胞表达。将在表达和不表达RoXaN的细胞中检查轮状病毒复制的效率、病毒和细胞信息的翻译以及病毒复制模式的变化。这些结果将确定RoXaN在轮状病毒复制中的作用。宿主实验室有独特的工具来检查这种迷人的新蛋白质的功能。宿主和他的同事是研究病毒操纵细胞生理学的专家，并且拥有完成该项目所需的资源。

0401866BeckerThe International Research Fellowship Program enables U.S. scientists and engineers to conduct three to twenty-four months of research abroad. The program's awards provide opportunities for joint research, and the use of unique or complementary facilities, expertise and experimental conditions abroad.This award will support a twelve-month research fellowship by Dr. Michelle M. Becker to work with Dr. Didier Poncet at Centre National de la Recherche Scientique in Gif-sur-Yvette, France.RoXaN I is a cellular protein with no known function, identified through its interaction with rotavirus nonstructural protein NSP3. RoXaN is found in both the nucleus and the cytoplasm and may move between these two cellular compartments, possibly to transport mRNA or regulate translation. These experiments are designed to test the hypothesis that the normal function of RoXaN is subverted during rotavirus infection to enhance viral replication and will also contribute new information about the mechanisms of rotavirus replication and viral control of cellular processes. Since RoXaN is present in both the cytoplasm and nucleus, but it does not contain a classical nuclear localization sequence or nuclear export sequence, the subcellular localization of RoXaN in uninfected and rotavirus-infected cells is being determined using confocal immunofluorescence microscopy. Deletion mutants of RoXaN are being used to define the protein domains necessary for proper subcellular localization. Results from these experiments will define the localization of RoXaN within the cell and the regions within the protein important for directing RoXaN to the correct cellular compartment. The effect of RoXaN on rotavirus processes will be examined in the presence and absence of RoXaN expression. Cellular expression of RoXaN mRNA is being deleted using RNA interference technology. The efficiency of rotavirus replication, translation of viral and cellular messages, and changes in viral replication patterns will be examined in cells expressing and not expressing RoXaN. These results will determine the role of RoXaN in rotavirus replication.The host lab has unique tools to examine the function of this fascinating and novel protein. The host and his colleagues are experts in studies of the viral manipulation of cellular physiology, and have the resources necessary to complete this project.