RUI: Genetic Analysis of Gurken translational Control in Drosophila

智力优势：所有动物都必须在不断变化的环境中重复建立自己的身体计划。Gurken是一种进化上保守的生长因子，在果蝇（Drosophila melanogaster）的卵子发生过程中，它被仔细调节以建立前/后和背/腹轴。该项目旨在通过解决Gurken mRNA对两种刺激的反应来确定Gurken翻译的调节机制：营养限制和DNA损伤。这项研究的第一个目标是了解果蝇在面临营养物质供应高度可变的环境时如何保持稳健的发育模式。蛋白质的生产受到高度调节，以应对营养的可用性，并确保资源的有效利用。当营养缺乏时，细胞中的翻译会受到全面抑制;然而，一小部分必需蛋白质通过采用称为IRES介导的翻译的替代机制来逃避这种抑制。PI实验室的初步工作表明，gurken mRNA利用这种替代机制来维持轴模式，当营养物质有限时。该研究将利用许多遗传和生物化学方法来研究gurken mRNA在体外和体内的活性。具体而言，将通过将gurken mRNA的调控区与发光荧光素酶报告基因连接来探测它们的活性。这将有助于发现IRES元件，使卵母细胞保持其强大的模式。研究的第二个目标是了解减数分裂过程中发生的基因组损伤如何影响gurken的翻译。先前的研究表明，干扰DNA修复的突变通过一种未知的机制抑制了gurken的翻译。已经分离出几种影响该信号通路的新突变。该项目将采用基因组测序的方法来识别这些突变，并揭示基因组完整性与发育模式分子之间的联系机制。更广泛的影响：该项目的研究将吸引州立大学纽约学院弗雷多尼亚分校的学生参与翻译调控研究，该学院是一所以本科为主的机构。如上所述，研究将解决该领域的几个紧迫问题，同时让学生成为活动背后的推动力。学生将密切参与实验的设计和执行，并有机会在区域和国家会议上介绍项目的结果。这项研究不仅会直接影响PI实验室的学生，还会影响核心遗传学实验室和高级分子遗传学实验室课程的学生（每年约70名学生）。该项目将促进三个暑期学生，将从弗雷多尼亚和/或区域社区学院招聘全职研究经验。

Intellectual merit: All animals must reproducibly establish their body plan in the face of a constantly changing environment. Gurken is an evolutionarily conserved growth factor that is carefully regulated to establish the anterior / posterior and dorsal / ventral axis during oogenesis in the fruit fly Drosophila melanogaster. This project aims to identify the mechanisms by which the translation of Gurken is regulated by addressing the response of gurken mRNA to two stimuli: Nutrient limitation and DNA-damage. The first goal of the research is to understand how Drosophila maintain robust developmental patterning when confronted with an environment where the availability of nutrients is highly variable. The production of proteins is highly regulated in response to nutrient availability and ensures the efficient utilization of resources. Translation in the cell is globally repressed when nutrients are scarce; however, a small subset of essential proteins escape this repression by employing an alternative mechanism known as IRES-mediated translation. Preliminary work from the PI's lab suggests that the gurken mRNA utilizes this alternative mechanism to maintain the axis patterning when nutrients are limiting. The research will utilize a number of genetic and biochemical approaches to study the activity of the gurken mRNA both in vitro and in vivo. Specifically, the activity of regulatory regions of the gurken mRNA will be probed by joining them to light emitting luciferase reporter genes. This will facilitate the discovery of IRES elements that allow the oocyte to maintain its robust patterning. The second goal of the research is to understand how damage to the genome that occurs during meiosis affects the translation of gurken. Previous research has shown that mutations that interfere with DNA repair inhibit the translation of gurken by an unknown mechanism. Several novel mutations that affect this signaling pathway have been isolated. This project will take a genomic sequencing approach to identify these mutations and shed light on the mechanism(s) that communicates the integrity of the genome to developmental patterning molecules.Broader Impact: The research conducted in this project will engage students in the study of translation regulation at the State University of New York College at Fredonia, a predominantly undergraduate institution. The research will address several pressing questions in the field, as indicated above, while allowing students to be the driving force behind the activities. The students will be intimately involved in the design and execution of the experiments and have the opportunity to present the results of the project at regional and national meetings. The research will directly impact not only the students working in the PI's laboratory, but also students that are enrolled in the core genetics lab and upper level molecular genetics lab courses (approximately 70 students per year). The project will facilitate full-time research experiences for three summer students that will be recruited from Fredonia and/or regional community colleges.