The Role of IME4 and IME2 in Drosophila Gametogenesis

描述(申请人提供)：我的研究的长期目标是阐明遗传程序中的早期事件，有助于利用果蝇确定后生动物的减数分裂细胞命运--这是后生动物的模式生物，我将在该奖项的指导阶段接受培训。我将从研究IME2和IME4开始我的研究，这是两个高度保守的基因，已被证明是酿酒酵母减数分裂所必需的。对决定减数分裂细胞命运的遗传程序的理解将为理解不孕不育和有丝分裂开关功能障碍的后果提供洞察力。IME2和IME4在小鼠、大鼠和人类中都有同源基因。具体目标：该建议的具体目标跨越两年的指导阶段和三年的独立阶段，在这两个阶段，我将培训黑腹葡萄球菌的畜牧学、遗传学、分子遗传学和细胞学技术，以研究IME4和IME2在减数分裂中的作用，在这三年的独立阶段，我将进一步追求我的发现，并确立自己作为独立研究人员的地位：第一阶段：目标1：通过经典遗传学和RNAi击倒，在黑腹果蝇中创建ime4和IME2的突变体，并启动它们的表型鉴定。目的：研究Ime4和IME2在卵子发生和精子发生中的特异性作用。第二阶段：目的3：研究Ime4作为一种RNA甲基转移酶在配子发生中的作用，并确定其修饰的靶点。目的4：确定IME2作为一种蛋白在配子发生中的作用，并确定其底物。与公共健康相关：男性和女性生殖细胞通过一种称为减数分裂的特殊细胞分裂，将其部分细胞的染色体含量减少一半，分别成为精子和卵子。决定减数分裂承诺的早期因素尚不清楚。我提议的实验旨在阐明使生殖细胞进入减数分裂的遗传程序。我的发现将为理解不孕不育和有丝分裂的适当功能的作用提供信息：减数分裂开关，其功能障碍在一些生殖系癌症中被怀疑。

DESCRIPTION (provided by applicant): The long-term goal of my studies is to elucidate the early events in the genetic program conducive to meiotic cell fate determination in metazoans using Drosophila melanogaster-the metazoan model organism in which I will be trained during the mentored phase of this award. I will start my research by investigating IME2 and IME4, two highly conserved genes that have been shown to be required for meiosis in S. cerevisiae. The understandig of the genetic program that determines meiotic cell fate will provide insight into understanding infertility and the consequences of dysfunction of the mitotic:meiotic switch. Both IME2 and IME4 have homologs in mice, rats, and humans. Specific aims: The specific aims of this proposal span both the two-year mentored phase of this award in which I will train in D. melanogaster's husbandry, genetics, molecular genetics and cytological techniques to investigate the role of IME4 and IME2 in meiosis, and the three year independent phase of this award in which I will further pursue my findings and establish myself as an independent investigator: Phase I: Aim 1: To create mutants of ime4 and ime2 in D. melanogaster by classical genetics and RNAi knock-downs and initiate their phenotypic characterization. Aim 2: To study the specific roles of Ime4 and Ime2 in oogenesis and spermatogenesis. Phase II: Aim 3: To investigate the role of Ime4 as an RNA methyltransferase in gametogenesis and identify its targets for this modification. Aim 4: To determine the role of Ime2 as a kinase in gametogenesis and identify its substrates. Relevance to public health: Male and female germ cells commit some of its cells to reduce by half their chromosomal content to become sperm and egg respectively by a specialized cell division called meiosis. The early factors that determine commitment to meiosis are not known. My proposed experiments are aimed at elucidatng the genetic program that makes a germ cell enter meiosis. My findings will provide information to understand infertility and the role of proper function of the mitosis:meiosis switch whose dysfunction has been suspected in some germline cancers.