SARCOMERIC PROTEINS IN NORMAL AND DISEASED MUSCLE

描述(逐字摘自调查人员摘要)：长期 这项建议的目标是识别和表征新的骨骼肌 肉瘤基因和蛋白质以及了解这些基因和蛋白质的异常 蛋白质有助于人类神经肌肉疾病的病理生理学。 此竞争性续订申请描述了以下补充方法 从基础科学和反向遗传学两方面解决这些问题 透视。第一个涉及六个新基因的继续表征。 在以骨骼肌α-肌动蛋白为基础的酵母双杂交筛选中鉴定 “诱饵。”对其中一种，现在被称为“myozenin”进行了广泛的描述， 发现它在许多体外生化分析中与α-肌动蛋白结合， 它共同定位于肌节的Z线。间接免疫荧光 线状肌病(NM)患者的肌肉分析显示 Myozenin异常地定位于线虫杆内。其他内容 Myozenin等新蛋白的生化和酵母双杂交研究 将提供对Z线和细丝结构和功能的新见解。 学习这些结构的一个补充方法是使用反转 识别和表征NM患者突变基因的遗传学 遗传性神经肌肉病的临床和遗传异质性组 以不同程度的进行性骨骼肌无力和 患病个体肌肉中特有的杆状小体。申请人的 实验室已在三个肉瘤基因中发现NM突变 (a-原肌球蛋白、星云蛋白和肌动蛋白)，并且有一个或 必须存在更多的NM基因。上面发现的每一个新的肉瘤基因， 以及一组已知的肉瘤候选基因，将被测试 NM基因突变。最后，目前还不清楚每个已知NM的突变是如何发生的 基因与极不稳定的临床和病理表型有关 在这种疾病中见过。为了解决这个发病机制的问题，全局基因 人NM骨骼肌的表达谱及转基因研究 NM小鼠模型将被开发出有趣的新的骨骼肌基因 谁的表情被扰乱了，将被研究。这个项目的成功将会 还可以对所有NM患者进行准确的诊断和预后测试 AS为正常和异常Z线的结构和功能提供了新的线索 和骨骼肌中的细丝。

DESCRIPTION (appended verbatim from investigator's abstract): The long term goals of this proposal are to identify and characterize new skeletal muscle sarcomeric genes and proteins and to understand how abnormalities of these proteins contribute to the pathophysiology of human neuromuscular diseases. This competitive renewal application describes complementary approaches to address these issues from both a basic science and reverse genetics perspective. The first involves continued characterization of six novel genes identified in a yeast two-hybrid screen using skeletal muscle a-actinins as "bait." Extensive characterization of one of these, now called "myozenin," reveals that it binds a-actinin in a number of in vitro biochemical assays and that it co-localizes at sarcomeric Z lines. Indirect immunofluorescence analysis of muscle from patients with nemaline myopathy (NM) reveals that myozenin is abnormally localized within the nemaline rods. Additional biochemical and yeast two-hybrid studies on myozenin and the other new proteins will provide new insights into Z line and thin filament structure and function. A complementary approach to learning about these structures is to use reverse genetics to identify and characterize the genes mutated in patients with NM, a clinically and genetically heterogeneous group of inherited neuromuscular diseases distinguished by variably progressive skeletal muscle weakness and characteristic rod bodies in muscles of affected individuals. The applicant's laboratory has identified NM mutations in three sarcomeric genes (a-tropomyosin, nebulin and actin) to date, and has genetic data that one or more additional NM genes must exist. Each new sarcomeric gene identified above, as well as a group of known sarcomeric candidate genes, will be tested for mutations in NM. Finally, it is not clear how mutations in each of the known NM genes relate to the extremely variable clinical and pathological phenotypes seen in this disease. To address this question of pathogenesis, global gene expression profiles of NM skeletal muscle from human patients and transgenic mouse models of NM will be developed and interesting new skeletal muscle genes whose expression is perturbed will be studied. Success in this project will enable accurate diagnostic and prognostic testing for all NM patients as well as shed new light on the structure and functions of normal and abnormal Z lines and thin filaments in skeletal muscle.