CELLULAR CONTROL OF NEURONAL CYTOSKELETON ASSEMBLY

拟议研究的长期目标是了解 神经母细胞瘤细胞的细胞骨架被组装和组织起来。角色 215000道尔顿微管相关蛋白(215K图)的 在神经母细胞瘤神经母细胞瘤细胞中存在，而在 未分化的细胞将被详细检查。活体分析将 建立该蛋白质在细胞各阶段的合成模式 附着和轴突形成。免疫荧光显微镜 针对215K图和微管蛋白的抗血清将指示分布 这些蛋白质在微管组织的不同阶段。这些 结果将与使用细胞分级获得的结果进行比较，以 分析了215K图相对于可溶性的分布和数量 和组装的微管蛋白。体外测试将通过以下方式建立机制 215K图谱与微管蛋白相互作用，生化程序将 证明这种蛋白质是否经过翻译后修饰。多肽 将使用测绘和克隆抗体来研究ExteNET 215K图与其他微管相关蛋白的同源性 在神经母细胞瘤细胞中，以及微管相关蛋白 其他细胞类型。将进行有限的研究，以检查 轴突形成过程中微管与神经丝的关系 并确定是否有任何微管与 蛋白质协调这两个细胞骨架系统。这些调查 应提供有关微管生物学作用的基本信息 微管组织中的相关蛋白，以及在 这类特殊蛋白质的生化性质。

The long term objective of the proposed research is to understand how the cytoskeleton of neuroblastoma cells is assembled and organized. The role of 215,000 dalton microtubule associated protein (215 K MAP) that is present in neurite-bearing neuroblastoma cells but absent in non-differentiated cells will be examined in detail. In vivo analyses will establish the synthetic pattern of this protein during stages of cell attachment and neurite formation. Immunofluorescence microscopy with antisera against the 215 K MAP and tubulin will indicate the distribution of these proteins at various stages of microtubule organization. These results will be compared to those obtained using cellular fractionation to analyze the distribtuion and amount of the 215 K MAP relative to soluble and assembled tubulin. In vitro assays will establish the mechanisms by which the 215 K MAP interacts with tubulin, and biochemical procedures will demonstrate whether this protein is post-translationally modified. Peptide mapping and monoclonal antibodies will be used to investigate the extenet of homology between the 215 K MAP and other microtubule associated proteins in neuroblastoma cells, as well as to microtubule associated proteins from other cell types. Limited studies will be undertaken to examine the relationship between microtubules and neurofilaments during neurite development, and to establish whether any of the microtubule associated proteins coordinate these two cytoskeletal systems. These investigations should provide basic information on the biological role of microtubule associated proteins in the organization of microtubules, and on the biochemical properties of this class of specialized proteins.