CELLULAR CONTROL OF CYTOSKELETON ASSEMBLY

这项拟议研究的长期目标是了解 不是马达的微管相关蛋白参与 微管的功能和组织。这些研究将集中在 MAP 4，一种微管相关蛋白，在早期表达 在小鼠的发育和特定的成体组织中。编码序列 已经获得了老鼠和人类的地图4，一个结构模型 定义的蛋白质和在横纹肌中观察到的新的转录本 睾丸。各种方法，包括转基因和/或 显微注射蛋白质片段或荧光衍生探针 将被应用于推断地图4的区域 函数，并询问是否存在与之交互的域 微管以外的细胞结构。肌源性细胞系，在 观察到肌肉特异性MAP 4转录本的诱导，威尔 被用来研究MAP4和微管蛋白的关系 肌管形成过程中同种异型的表达和分布。反义 技术将被用来询问MAP 4是否对正常 细胞功能，或诸如肌肉发生或植入前的事件 发展。MAP 4在小鼠肌肉发生、精子发生过程中的表达 早期发育将通过相关的免疫细胞学进行研究 和原位杂交分析。进一步的分子表征 将进行编码序列的编码以确定数目， 多个mRNAs的相互关系和产生机制 可能会产生多种蛋白质异构体。一个小项目将会 是表明RII亚基的实验的完成 CAMP依赖的蛋白激酶介导MAP-2与 高尔基。

The long range goal of the proposed research is to understand how microtubule-associated proteins that are not motors participate in the function and organization of microtubules. These studies will focus on MAP 4, a microtubule-associated protein that is expressed during early development and in specific adult tissues in mice. Coding sequences for mouse and human MAP 4 have been obtained, a structural model for the protein defined, and novel transcripts observed in striated muscle and testis. A variety of approaches, including transfection and/or microinjection of protein fragments or fluorescently derivatized probes will be applied to deduce the regions of MAP 4 that are essential for function, and to ask whether or not there are domains that interact with cellular structures other than microtubules. Myogenic cell lines, in which induction of muscle-specific MAP 4 transcripts is observed, will be employed to examine the relationship between MAP 4 and tubulin isotype expression and distribution during myotube formation. Antisense technology will be used to ask whether MAP 4 is essential for normal cellular function, or events such as myogenesis or pre-implantation development. MAP 4 expression during mouse myogenesis, spermatogenesis and early development will be studied with correlative immunocytological and in situ hybridization analyses. Further molecular characterization of the coding sequences will be undertaken to determine the number, relationship, and mechanism of generation of the multiple MRNAS which presumably give rise to multiple protein isoforms. A minor project will be the completion of experiments that indicate the RII subunit of CAMP-dependent protein kinase mediates interaction of MAP 2 with the Golgi.