Collaborative Proposal to Examine the Function and

检查功能和功能的协作提案

• 批准号: 1456023
• 负责人: Leonard Dobens
• 金额: $ 82万
• 依托单位: University of Missouri-Kansas City
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1456023&HistoricalAwards=false
• 关键词: Collaborative; Proposal; Examine; Function

A fundamental problem in biology is how the proper size of an organism is achieved. The size of tissues and organs is a consequence of both cell size and number and is controlled by both extrinsic signals, such as insulin peptides, and intrinsic cell signals that measure nutritional and environmental inputs. This research program will exploit a genetic model organism to understand how cell division is balanced by cell growth by the action of a gene named Tribbles. Tribbles is found in all animals, and in humans it has been connected to cancer and Type 2 diabetes, but its functions are poorly understood. It has been shown that Tribbles blocks: (1) cell division by binding the protein Cdc25, a key trigger of division, and (2) insulin-stimulated cell growth by binding the protein Akt kinase. This research program will shed light on the functions of Tribbles that regulate animal size during development, and will have implications for scientists studying the role of Tribbles in human disease. The research program will also support ongoing efforts to train students in molecular and cell biological approaches to dissect conserved developmental mechanisms in a model organism.These studies of the conserved role of Trbl in cell growth and proliferation will offer insight into the genetic basis of animal diversity, the mechanism of evolution, the effect of environmental conditions on a developmental program, and the underlying causes of developmental aberrations and metabolic disease. Body sizes vary impressively between closely related animal species, and species-specific adaptations, such as the mammalian forelimb and specialized head segments among insects, show incredible size scaling. Size is subject to intense evolutionary selection pressure with consequences for mate selection, predation and tolerance to environmental changes, moreover animals adjust size to both nutrient availability and a host of environmental cues including temperature and O2 levels. The mechanisms coordinating this developmental plasticity are incompletely understood, and include integration of cues from hormonal signals, growth factors and insulin-like peptides to ensure that energy expended on growth matches energy intake. The kinase Tribbles (Trbl) binds and blocks a number of key targets that regulate cell growth, division and differentiation, notably cdc25 phosphatase to inhibit cell proliferation and Akt kinase to inhibit insulin-stimulated cell growth and metabolism. The notion that Trbl is an conserved adaptor protein that regulates both cell proliferation (by binding and degrading Cdc25) and cell size (by binding and inhibiting Akt) in response to diet will be tested in three aims: (1) to explore the role of Trbl in cell growth during normal and dietary stress; (2) to perform a structure/function analysis of Trbl, focused on its conserved features; and (3) to conduct a screen for proteins that physically interact with Trbl by (a) yeast two hybrid screens and (b) co-immuno-pulldown using Trbl-specific antisera.

生物学中的一个基本问题是如何获得有机体的适当大小。组织和器官的大小是细胞大小和数量的结果，并且由外在信号（例如胰岛素肽）和测量营养和环境输入的内在细胞信号控制。这项研究计划将利用一种遗传模式生物来了解细胞分裂是如何通过一种名为Tribbles的基因的作用来平衡细胞生长的。Tribbles存在于所有动物中，在人类中，它与癌症和2型糖尿病有关，但人们对其功能知之甚少。已经表明，Tribbles阻断：（1）通过结合蛋白Cdc 25（分裂的关键触发物）的细胞分裂，和（2）通过结合蛋白Akt激酶的胰岛素刺激的细胞生长。 这项研究计划将揭示Tribbles在发育过程中调节动物大小的功能，并将对研究Tribbles在人类疾病中作用的科学家产生影响。该研究计划还将支持正在进行的努力，培养学生在分子和细胞生物学方法，剖析保守的发展机制，在一个模式生物。这些研究的保守作用Trbl在细胞生长和增殖将提供深入了解动物多样性的遗传基础，进化机制，环境条件对发育程序的影响，以及发育异常和代谢疾病的根本原因。 在密切相关的动物物种之间，身体大小的变化令人印象深刻，物种特异性的适应，如哺乳动物的前肢和昆虫中专门的头节，显示出令人难以置信的大小缩放。大小受到强烈的进化选择压力，对配偶选择，捕食和对环境变化的耐受性产生影响，此外，动物会根据营养物质的可用性和许多环境线索（包括温度和O2水平）来调整大小。协调这种发育可塑性的机制还不完全清楚，包括整合来自激素信号、生长因子和胰岛素样肽的线索，以确保生长所消耗的能量与能量摄入相匹配。激酶Tribbles（Trbl）结合并阻断许多调节细胞生长、分裂和分化的关键靶标，特别是cdc 25磷酸酶以抑制细胞增殖和Akt激酶以抑制胰岛素刺激的细胞生长和代谢。Trbl是一种保守的衔接蛋白，调节细胞增殖和细胞增殖，（通过结合和降解Cdc 25）和细胞大小（通过结合和抑制Akt）对饮食的反应将在三个目标中进行测试：（1）探索Trbl在正常和饮食应激期间细胞生长中的作用;（2）进行Trbl的结构/功能分析，集中于其保守特征;和（3）通过（a）酵母双杂交筛选和（B）使用Trbl-特异性抗血清的共免疫-下拉来筛选与Trbl物理相互作用的蛋白质。