CELLULAR CONTROL OF CYTOSKELETON ASSEMBLY

拟议研究的长期目标是了解如何 不是马达的微管相关蛋白参与了 微管的功能和组织。 这些研究将侧重于 MAP 4是一种微管相关蛋白，在早期表达。 发育和小鼠特定的成年组织中。 的编码序列 小鼠和人的MAP 4已经获得， 蛋白质定义，并在横纹肌和 睾丸 多种方法，包括转染和/或转染， 蛋白质片段或荧光衍生探针的显微注射 将被应用于推导MAP 4的区域，这些区域对于 函数，并询问是否存在与 除了微管之外的细胞结构。 生肌细胞系， 观察到肌肉特异性MAP 4转录物的诱导， 探讨MAP 4与微管蛋白的关系 肌管形成过程中同种型表达和分布。 反义 技术将被用来询问MAP 4是否对正常的 细胞功能，或事件，如肌生成或植入前 发展 MAP 4在小鼠肌肉发生、精子发生中的表达 并对早期发育进行相关免疫细胞学研究 和原位杂交分析。 进一步分子表征 来确定编码序列的数目， 的关系，以及多种MRNAS的产生机制， 推测产生多种蛋白质同种型。 一个小项目将 完成实验表明，RII亚基的 cAMP依赖性蛋白激酶介导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.