DROSOPHILA GENES AFFECTING CHROMOSOME SEGREGATION

这项建议的重点是遗传和分子检测。 果蝇的zw10基因。Zw10基因中的突变破坏了 有丝分裂细胞和雄性减数分裂细胞中的染色体分离。 人们对ZW10蛋白的兴趣很大程度上源于它的动态 有丝分裂过程中的定位。私家侦探和他的合作者辩称 这种蛋白可能在调节动粒的过程中起着关键作用 功能，并在发送信号或响应存在或 动粒上没有张力。Zw10基因编码一种 85kd的蛋白，与其他蛋白没有明显的同源性 已知功能。PI提出了一系列巧妙的实验 旨在阐明这种蛋白质的功能。具体来说，他 建议(1)使用免疫EM和RT视频显微镜来 高空跟踪ZW10的时空分布 决议；(2)调查注射抗ZW10的后果 不同时间的抗体；(3)测定对ZW10的影响 激光手术对染色体张力变化的影响 并通过微操作；(4)确定修改是否 的ZW10蛋白(例如，磷酸化)与其 细胞周期中的运动；(5)执行结构/功能 (6)对ZW10蛋白的功能进行了研究。 (7)研究ZW10与线虫的关系。 其他已知基因的产物；(8)识别相互作用的蛋白质 用ZW10目前标准的生化和遗传手段； (9)开始对一种新基因进行分子分析，SAN，其 蛋白质产物似乎在姐妹中扮演着重要的角色 染色单体凝聚力。

This proposal is focused on the genetic and molecular examination of the Drosophila zw10 gene. Mutants in the zw10 gene disrupt chromosome segregation in both mitotic and male meiotic cells. Much of the interest in the ZW10 protein stems from its dynamic localization during mitosis. The PI and his collaborators argue that this protein may play a critical role in mediating kinetochore function and in signaling, or responding to, the presence or absence of tension on the kinetochore. The zw10 gene encodes an 85 kd protein with no significant homologies to other proteins of known function. The PI proposes a clever set of experiments designed to elucidate the function of this protein. Specifically, he proposes to (1) use immuno-EM and RT videomicroscopy to follow the temporal and spatial distribute of ZW10 at very high resolution; (2) investigate the consequences of injecting anti-ZW10 antibodies at various times; (3) assay the effects on ZW10 distribution of altering tension on chromosomes by Iaser surgery and by micro-manipulation; (4) determine whether modifications of the ZW10 protein (e.g. phosphorylation) are associated with its movements during the cell cycle; (5) perform a structure/function dissection of ZW10; (6) study the function of the ZW10 protein in C elegans.; (7) investigate the relationship of ZW10 to the products of other known genes; (8) identify proteins that interact with ZW10 by now standard biochemical and genetic means; and (9) begin the molecular analysis of a new gene, san, whose protein product appears to play an essential role in sister chromatid cohesion.