CONTROL OF MEIOSIS AND GERMLINE PROLIFERATION

生殖系是有性繁殖所必需的永生细胞谱系。 繁殖。生殖系的细胞周期可能会非常不同 在发育调节方面有别于体细胞， 检查点和细胞分裂机制。我们在C. 优雅的人已经确定emb-30基因座是一个潜在的生殖系- 生殖细胞增殖所需的特定细胞周期基因， 卵母细胞和精母细胞完成减数分裂。 我们的分析表明，Emb-30卵母细胞在退出时存在缺陷， 但没有进入减数分裂中期：受精后，它们 不能进展到减数分裂后期I和一个无序的停滞 纺锤形。所有后续的胚胎事件都被阻止了。此外, Emb-30似乎在生殖系增殖和 配子队形。重要的是，所有已知的emb-30突变表型都是 是生殖系特有的，没有细胞分裂或缺陷 在任何体细胞中都观察到了分化。基于 反义RNA表型复制结果和遗传作图结果，emb-30可以 编码一种与SRPK1蛋白激酶相关的蛋白，该蛋白和 磷酸化剪接蛋白并影响其核 分隔化。一种模式是emb-30扮演着关键角色。 控制生殖系中细胞周期调节因子的剪接。 或者，emb-30可能直接影响生殖系细胞周期， 独立于剪接，可能通过影响减数分裂纺锤体，或一个 主轴检查点。为了确定emb-30在生殖系中的作用 发展，我们将检验以下假设：1)EMB-30是 减数分裂和生殖系增殖所必需的；2) Emb-30编码与SRPK1相关的保守蛋白激酶；和3) Emb-30在基因上与GLD-1和GLP-1相互作用，这两个基因 已经被明确地证明在生殖系发育中发挥作用。按顺序 为了检验这些假设，我们将：a)分离和表征新的emb- 40个等位基因；b)对emb-30进行广泛的分子分析；c) 分析与GLD-1和GLP-1的上位性交互作用。建议数 研究将加深我们对减数分裂细胞周期的理解，并可能 统一我们对生殖系中有丝分裂和减数分裂的看法 共享基本组件。鉴于它们在生物学上的广泛相似性 在远缘生物之间的减数分裂过程中， 对生殖系发育的进化限制，这项研究是 可能为减数分裂的控制提供重要的新见解 在后生动物中。从这些研究中获得的信息将是有用的 了解人类生育/不育以治疗生殖系 肿瘤。

The germline is an immortal cell lineage essential for sexual reproduction. The germline cell cycle is likely to be very different from that of somatic cells in terms of developmental regulation, checkpoints, and cell division mechanisms. Our genetic studies in C. elegans have identified the emb-30 locus as a potential germline- specific cell cycle gene, required both for germ cells to proliferate, and for oocytes and spermatocytes to complete their meiotic divisions. Our analysis suggests that emb-30 oocytes are defective in exit from, but not entry into, meiotic metaphase: Following fertilization, they cannot progress to anaphase of meiosis I and arrest with a disorganized spindle. All subsequent embryonic events are blocked. In addition, emb-30 appears to play an essential role in germline proliferation and gamete formation. Importantly, all known emb-30 mutant phenotypes are specific to the germline and no defects in cell division or differentiation have been observed in any somatic cell. Based on antisense RNA phenocopy results, and genetic mapping results, emb-30 may encode a protein related to the SRPK1 protein kinase, which and phosphorylate splicing proteins and affect their nuclear compartmentalization. One model is that emb-30 plays a critical role in controlling the splicing of cell cycle regulators in the germline. Alternatively, emb-30 may directly affect the germline cell cycle, independent of splicing perhaps by affecting the meiotic spindle, or a spindle checkpoint. In order to define the role of emb-30 in germline development, we will test the following hypotheses: 1) that emb-30 is required specifically for meiosis and germline proliferation; 2) that emb-30 encodes a conserved protein kinase related to SRPK1; and 3) that emb-30 genetically interacts with gld-1 and glp-1, two genes that have been shown definitively to function in germline development. In order to test these hypotheses, we will: a) isolate and characterize new emb- 40 alleles; b) conduct an extensive molecular analysis of emb-30; c) analyze epistatic interactions with gld-1 and glp-1. The proposed studies will deepen our understanding of the meiotic cell cycle and may unify our view of mitosis and meiosis in the germline by identifying a shared essential component. Given the extensive biological similarities in the meiotic process between distantly related organisms, and evolutionary constraints on germline development, this research is likely to provide significant new insights into the control of meiosis in metazoans. The information gained from these studies will be useful for understanding human fertility/infertility for treating germline tumors.