Super-resolution microscopy of nuclear lamin and spindle envelope/matrix function

描述（由申请人提供）：本提案旨在使用超分辨率显微镜技术研究间期和有丝分裂期间不同类型的层蛋白网络/网状结构的结构组织。在Aim 1中，我们计划确定每个层状蛋白（LA， LC, LB1, LB2）是否形成单独的层状网，或者层状网的建立是否需要正常间期细胞核中不同层状蛋白同种异构体的相互作用。我们还将研究每个层状蛋白与染色质和核孔复合物（NPCs）的相互作用。当动物细胞进入有丝分裂时，包括核层在内的部分间期膜网络和核蛋白重新组织成一个相互连接的网络，包围并渗透纺锤体微管。这个网络被称为纺锤包络（或鞘）或纺锤矩阵。这里我们将网络称为主轴包络/矩阵（Spenix）。定义Spenix功能的主要挑战是可视化Spenix蛋白的行为，并将其行为与有丝分裂中的功能联系起来。在Aim 2中，我们将使用超分辨率显微镜研究Spenix的膜和蛋白质组织，这将为解剖Spenix的组织以及Spenix与有丝分裂纺锤体的功能关系打开大门。所提出的研究结果将为细胞核层板的相互作用和功能及其在间期和有丝分裂过程中与主要相互作用伙伴的相互作用提供重要的新信息。

DESCRIPTION (provided by applicant): This proposal aims to study the structural organization of the different types of lamin networks/meshworks during interphase and mitosis using super resolution microscopy techniques. In Aim 1, we plan to determine whether each lamin (LA, LC, LB1, LB2) forms a separate lamina meshwork or whether the establishment of a lamina meshwork requires the interactions of different lamin isoforms in normal interphase nuclei. We will also study the interactions of each of the lamins with chromatin and nuclear pore complexes (NPCs). As animal cells enter mitosis, parts of the interphase membrane network, including the nuclear lamina, and nuclear proteins re-organize into an inter-connected network that both surrounds and permeates the spindle microtubules. This network has been referred to as either the spindle envelope (or sheath) or spindle matrix. Here we will refer to the network as the Spindle envelope/matrix (Spenix). The major challenge in defining the function of the Spenix is to visualize the behavior of Spenix proteins and link their behaviors to their functions in mitosis. In Aim 2, we will apply super resolution microscopy to study the membrane and protein organization of the Spenix, which should open the door to dissect not only the organization of the Spenix but also the functional relationship of the Spenix and the mitotic spindle. The results from the proposed studies will provide important new information regarding the interactions and functions of the nuclear lamins and their interactions with their major interacting partners in interphase and mitosis.