Functional Study of Native and Synthetic Nuclei

描述（由申请人提供）：细胞核和细胞质之间的交通必须通过复杂和高度调控的核孔复合物，因此作为基因组的门户。正常交通包括来自外部世界的所有信号，这些信号被改写成一种唤起核变化的语言。对外交通包括旨在将这种核变化转化为行动的信息。一项疾病研究表明，包括HIV、流感、脊髓灰质炎和SV40在内的病毒已经进化出了直接攻击核孔和核运输中心的机制。遗传性人类疾病，如与Nup98和nup214相关的人类白血病，是由这些核孔蛋白基因的破坏造成的。要了解疾病和病毒感染是如何从破坏正常的核贩运中产生的，就必须首先了解这种贩运的机制。最近，我们的实验室发现了一个特定的亚基，Nup 107-160亚复合物，它对核孔的结构、功能和组装至关重要。如果没有这个亚复合体，细胞核就没有核孔。目前的资助有四个主要目标：(1)首先是获得关键的Nup107-160复合物的完整分子和结构知识。这些知识将用于破译核孔内复杂的相互作用，从而可以建立复杂的脊椎动物核孔的第一个三维地图。(2)第二个目标是了解核孔组装的机制，这是每个细胞周期必须发生的关键事件。Nup107-160复合体对NPC大会的本质为这个任务提供了一个很好的起点。(3)第三个目标是定义一个意想不到的玩家（输入beta）如何不仅调节NPC组装，还控制核膜融合和组装。将寻求在这些监管角色中引入β的目标。(4)最后一个目标是确定Nup107- 160复合体在有丝分裂着丝点中的功能。成功的完成应该提供对通往核心的重要通道的基本洞察。

DESCRIPTION (provided by applicant): Traffic between the nucleus and cytoplasm must pass through the elaborate and highly regulated nuclear pore complex, which thus serves as a gateway to the genome. Normal traffic includes all signals from the outside world, reworded into a language that evokes nuclear change. Traffic outward includes messages designed to translate this nuclear change into action. A study of disease reveals that viruses, including HIV, influenza, polio, and SV40, have evolved mechanisms to hit right at the heart of the nuclear pore and nuclear transport. Inherited human diseases, such as the Nup98- and Nup214-linked human leukemias, result from disruption of these nucleoporin genes. To understand the ways in which disease and viral infection arise from the subversion of normal nuclear trafficking, one must first understand the mechanism of that trafficking. Recently, our lab identified a specific subunit, the Nup 107-160 subcomplex, which is essential to the structure, function, and assembly of the nuclear pore. In the absence of this subcomplex, nuclei are devoid of nuclear pores. The present grant has four major goals: (1) The first is to gain a full molecular and structural knowledge of the critical Nup107-160 complex. This knowledge will be used to decipher complex interactions within the nuclear pore, so that the first three-dimensional map of the elaborate vertebrate nuclear pore may be built. (2) A second goal is to understand the mechanism of nuclear pore assembly, a key event that must occur each cell cycle. The essential nature of the Nup107-160 complex to NPC assembly gives an excellent starting point for this quest. (3) A third goal is to define how an unexpected player, importin beta, not only regulates NPC assembly, but also controls nuclear membrane fusion and assembly. The targets of importin beta in each of these regulatory roles will be sought. (4) A last goal focuses on determining the function of the Nup107- 160 complex on mitotic kinetochores. Successful completion should provide essential insight into the vital gateway to the nucleus.