MOLECULAR GENETICS OF PRONUCLEAR FUNCTIONS IN DROSOPHILA

果蝇原核功能的分子遗传学

• 批准号: 3303893
• 负责人: Mariana Federica Wolfner
• 金额: $ 15.54万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3303893
• 关键词: DNA binding protein; Drosophilidae; autoradiography; cell cycle; early embryonic stage; electrofocusing; electron microscopy; embryo /fetus protein; gene expression; gene interaction; gene mutation; genetic regulation; glycoproteins; in situ hybridization; macromolecule; molecular genetics; nonmammalian vertebrate embryology; nuclear membrane; nucleic acid sequence; polymerase chain reaction; protein sequence; protein structure function; site directed mutagenesis; spectrometry

The uniting of haploid chromosome sets from an egg and a sperm initiates the formation of a new diploid organism and triggers the onset of mitosis in embryos. I propose to analyze parentally-encoded gene products needed for these very early embryonic events in Drosophila. Specifically, we will continue our molecular genetic analysis of the fs(1)Ya gene, and extend our studies to gene products that interact with or control it. Function of the fs(1)Ya gene is required for the very first (gonomeric) division and possibly for subsequent mitotic divisions in early embryos. The fs(1)Ya gene encodes a maternally-specified nuclear envelope protein essential for the integrity for the nuclear lamina. The protein's association with the nuclear envelope is cell cycle stage dependent. Our studies will address the role and interactions of parental macromolecules, and of the nuclear envelope, in early embryonic mitosis. We will complete the characterization of embryos produced by fs(1)Ya mutant mothers by examining nuclear envelopes and mitotic components in these embryos. Then we will investigate biochemical characteristics suggested by the sequence of the fs(1)Ya protein and the phenotypes of fs(1)Ya null mutants. These include: whether the fs(1)Ya protein can be phosphorylated, whether it can bind to lamins and whether it can bind to DNA. The sequence alterations in fs(1)Ya mutations will be determined and correlated with the subcellular distribution of the mutant proteins and the mutant phenotypes to determine functional domains of the protein. In order to place fs(1)Ya action in a temporal order with the action of other parental gene products controlling mitosis, we will continue to examine the phenotypes of double mutants of fs(1)Ya2 and "early arrest" mutations. We will also examine the fs(1)Ya protein in embryos of mutants in genes that function in early mitoses. To identify genes whose products directly interact with fs(1)Ya protein, we will screen for dominant suppressors of fs(1)Ya mutations. In future years, the genes identified in the suppressor screens will be analyzed and the nature of the interactions between their products and fs(1)Ya protein determined. The proposed studies will allow a molecular genetic dissection of macromolecules needed for pronuclear fusion and initiation of mitosis in embryos. Simultaneously, they will permit the first genetic dissection of a non-lamin nuclear envelope component, and its developmental function, in an animal.

卵子和精子的单倍体染色体组的结合开始 形成新的二倍体生物体并触发有丝分裂的开始 在胚胎中。 我建议分析所需的亲本编码基因产物 对于果蝇中这些非常早期的胚胎事件。 具体来说，我们将 继续我们对 fs(1)Ya 基因的分子遗传学分析，并扩展我们的研究 研究与其相互作用或控制它的基因产物。 的功能 fs(1)Ya 基因是第一次（生殖）分裂所必需的，并且 可能用于早期胚胎随后的有丝分裂。 fs(1)Ya 基因编码母系特异的核膜蛋白，该蛋白对于 核层的完整性。 该蛋白质与 核膜是细胞周期阶段依赖性的。 我们的研究将解决 亲本大分子和核的作用和相互作用 早期胚胎有丝分裂中的包膜。 我们将完成 fs(1)Ya 突变体产生的胚胎的表征 母亲通过检查这些核膜和有丝分裂成分 胚胎。 然后我们将研究以下建议的生化特征 fs(1)Ya 蛋白的序列和 fs(1)Ya 的表型 null 突变体。 其中包括：fs(1)Ya蛋白是否可以被磷酸化， 是否可以与核纤层蛋白结合以及是否可以与DNA结合。 顺序 fs(1)Ya 突变的改变将被确定并与 突变蛋白的亚细胞分布和突变表型 以确定蛋白质的功能域。 为了将 fs(1)Ya 动作按时间顺序排列为 其他控制有丝分裂的亲本基因产物，我们将继续 检查 fs(1)Ya2 和“早期逮捕”双突变体的表型 突变。 我们还将检查突变体胚胎中的 fs(1)Ya 蛋白 在早期有丝分裂中起作用的基因中。 识别其产物的基因 直接与 fs(1)Ya 蛋白相互作用，我们将筛选显性的 fs(1)Ya 突变的抑制因子。 在未来的几年里，在 将分析抑制器屏幕和相互作用的性质 他们的产品和 fs(1)Ya 蛋白之间的测定。 拟议的研究将允许对 原核融合和有丝分裂启动所需的大分子 胚胎。 同时，他们将允许对 非核纤层核膜成分及其发育功能 一只动物。