The Role of IME4 and IME2 in Drosophila Gametogenesis

描述（由申请人提供）：我研究的长期目标是利用果蝇（Drosophila melanogaster）阐明有助于决定后生动物减数分裂细胞命运的遗传程序的早期事件——我将在本奖项的指导阶段接受后生动物模型生物的培训。我将从研究IME2和IME4开始我的研究，这两个高度保守的基因已经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症和有丝分裂减数分裂开关功能障碍的后果提供见解。IME2和IME4在小鼠、大鼠和人类中都有同源物。具体目标：该提案的具体目标包括两年的指导阶段，在此期间我将接受D. melanogaster的饲养，遗传学，分子遗传学和细胞学技术的培训，以研究IME4和IME2在减数分裂中的作用，以及三年的独立阶段，在此期间我将进一步追求我的发现并建立自己作为独立研究者的身份。通过经典遗传学和RNAi敲除，在黑腹龙骨中产生ime4和ime2突变体，并启动其表型表征。目的2：研究Ime4和Ime2在卵子发生和精子发生中的具体作用。II期：目的3：研究Ime4作为RNA甲基转移酶在配子体发生中的作用，并确定其修饰的靶标。目的4：确定Ime2作为一种激酶在配子体发生中的作用，并确定其底物。与公共卫生的相关性：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，使其一些细胞的染色体含量减少一半，分别成为精子和卵子。决定减数分裂的早期因素尚不清楚。我提出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症和有丝分裂正常功能的作用提供信息：减数分裂开关的功能障碍已被怀疑在一些种系癌症中。

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.