CAREER: Reorganization of the Vertebrate Golgi Apparatus
职业:脊椎动物高尔基体的重组
基本信息
- 批准号:9875939
- 负责人:
- 金额:$ 50万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:1999
- 资助国家:美国
- 起止时间:1999-03-01 至 2004-02-29
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The Golgi apparatus is organized into stacks of membrane-enclosed cisternae, but the processes that generate this remarkable structure are still poorly understood. Conventional models represent the Golgi as a series of stable, well-defined subcompartments. However, emerging data favor a 'cisternal maturation' model that views the Golgi not as an independent organelle, but as a dynamic outgrowth of the endoplasmic reticulum. Golgi cisternae are postulated to form de novo by the fusion of ER-derived vesicles, which are produced at specialized subdomains called transitional ER (tER) sites.The goal of this research is to understand the processes that create tER sites and Golgi stacks. As part of this effort, the current proposal focuses on characterizing the mechanisms that mediate reorganization of the Golgi apparatus. The Golgi of vertebrate cells undergoes dramatic rearrangements under certain conditions, including depolymerization of microtubules and passage of the cells through mitosis. Based on the cisternal maturation model, it seems plausible that some of these reorganization events involve the transit of Golgi components through the ER, followed by the emergence of new Golgi structures next to tER sites. This hypothesis will be tested by a combination of biochemical assays and fluorescence microscopy studies.The research portion of this proposal has two Specific Aims:1. TO DEVELOP A BIOCHEMICAL ASSAY THAT WILL REVEAL WHETHER GOLGI PROTEINS TRANSIT THROUGH THE ER DURING GOLGI REORGANIZATION. Transit through the ER will be detected by the action of ER-localized glucosidases. These glucosidases will initially be blocked with specific inhibitors, thereby loading the Golgi with proteins that contain glucosylated oligosaccharides. After removal of the inhibitor, Golgi reorganization will be induced, and then Golgi proteins will be analyzed for the presence or absence of glucose residues.2. TO ANALYZE THE LINK BETWEEN tER FUNCTION AND GOLGI REORGANIZATION USING FLUORESCENCE MICROSCOPY. The cisternal maturation model predicts that the distribution and activity of tER sites will influence Golgi reorganization. This prediction will be tested using fluorescence microscopy. For example, if a drug treatment initially causes Golgi proteins to return to the ER, the reorganized Golgi structures should be found near tER sites. To examine the dynamics of tER sites, these sites will be labeled with a Green Fluorescent Protein (GFP) fusion construct.These experiments will synergize with ongoing analysis of budding yeasts in the Glick laboratory. By elucidating conserved aspects of the tER-Golgi system, these studies aim to provide a unified molecular description of Golgi stack formation.This research plan is closely integrated with Dr. Glick's CAREER education objectives. In his undergraduate and graduate cell biology classes, the Golgi serves as a useful example for illustrating three key lessons: 1, cell biological processes are best understood by synthesizing data from diverse model organisms; 2, cells were designed not by a logical engineer, but by an opportunistic evolution; and 3, one must always be willing to reconsider basic assumptions. More generally, a recurring theme from Dr. Glick's explorations of the Golgi is the importance of viewing each question from multiple perspectives. This point is emphasized strongly in Dr. Glick's teaching, which focuses on developing students' capacity for critical thinking. In turn, the exercise of formulating and presenting these insights stimulates the teacher's own thinking about research-related questions.
高尔基体被组织成一堆堆膜封闭的池,但产生这种显着结构的过程仍然知之甚少。传统的模型表示高尔基体作为一系列稳定的,明确的亚室。然而,新出现的数据有利于“池成熟”模型,认为高尔基体不是一个独立的细胞器,但作为一个动态的产物的内质网。高尔基体池被认为是由内质网衍生的囊泡融合而形成的,这些囊泡产生于称为过渡性内质网(tER)位点的专门亚域。作为这项工作的一部分,目前的建议侧重于表征的机制,介导重组的高尔基体。脊椎动物细胞的高尔基体在某些条件下经历戏剧性的重排,包括微管的解聚和细胞通过有丝分裂。基于脑池成熟模型,似乎有道理的是,这些重组事件中的一些涉及高尔基体组件通过ER的过境,随后出现新的高尔基体结构旁边的tER网站。这个假设将通过生化分析和荧光显微镜研究相结合的方法来检验。进行一项生化检测,以揭示高尔基体重组期间高尔基体蛋白是否通过ER转运。通过ER的转运将通过ER定位的葡糖苷酶的作用来检测。这些葡糖基化酶最初会被特定的抑制剂阻断,从而使高尔基体负载含有葡糖基化寡糖的蛋白质。去除抑制剂后,将诱导高尔基重组,然后分析高尔基蛋白是否存在葡萄糖残基。2.用荧光显微镜分析tER功能与Golgi重组的关系。脑池成熟模型预测tER位点的分布和活性将影响高尔基体的重组。这一预测将使用荧光显微镜进行测试。例如,如果药物治疗最初导致高尔基体蛋白返回ER,那么重组的高尔基体结构应该在tER位点附近找到。为了检测tER位点的动力学,这些位点将用绿色荧光蛋白(GFP)融合构建体标记。这些实验将与Glick实验室正在进行的芽殖酵母分析协同作用。通过阐明tER-高尔基体系统的保守方面,这些研究旨在提供一个统一的高尔基体堆叠形成的分子描述。这项研究计划与Glick博士的职业教育目标紧密结合。在他的本科生和研究生细胞生物学课程中,高尔基体是一个有用的例子,说明了三个关键的教训:1,细胞生物学过程最好通过综合来自不同模式生物的数据来理解; 2,细胞不是由逻辑工程师设计的,而是由机会主义进化而来的; 3,人们必须总是愿意重新考虑基本假设。 更一般地说,格利克博士对高尔基体的探索中反复出现的一个主题是从多个角度看待每个问题的重要性。这一点在格利克博士的教学中得到了强烈的强调,他的教学重点是培养学生的批判性思维能力。反过来,制定和提出这些见解的练习刺激了教师自己对研究相关问题的思考。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Benjamin Glick其他文献
RDARuntime: An OS for AI Accelerators
RDARuntime:人工智能加速器操作系统
- DOI:
- 发表时间:
2023 - 期刊:
- 影响因子:0
- 作者:
Benjamin Glick;Arjun Sabnis;Renate Kempf;Arnav Goel;Aarti Lalwani;Guoyao Feng;Kiran Ranganath - 通讯作者:
Kiran Ranganath
Pneumopericardium during continuous positive airway pressure in respiratory distress syndrome.
呼吸窘迫综合征持续气道正压通气期间的心包积气。
- DOI:
10.1097/00003246-198412000-00018 - 发表时间:
1984 - 期刊:
- 影响因子:8.8
- 作者:
AI. PAN Gad;Kami Coder;Benjamin Glick;Ofra Peleg;A. Avital;Fabian Eyal - 通讯作者:
Fabian Eyal
Benjamin Glick的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Benjamin Glick', 18)}}的其他基金
Acquistion of a High-Pressure Freezing/Freeze Substition System for Electron Tomography and Immunocytochemistry
获得用于电子断层扫描和免疫细胞化学的高压冷冻/冷冻替代系统
- 批准号:
0420958 - 财政年份:2004
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
A DIC/Fluorescence Microscope for the Cummings Life Science Center
卡明斯生命科学中心的 DIC/荧光显微镜
- 批准号:
9970185 - 财政年份:1999
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Analysis of the Inheritance and Positioning of the Golgi Apparatus
高尔基体的传承与定位分析
- 批准号:
9604342 - 财政年份:1997
- 资助金额:
$ 50万 - 项目类别:
Continuing Grant
相似海外基金
Systematic Vertebrate Collections in Archaeological Research: Stabilization and Reorganization of The University of Tennessee Comparative Vertebrate Collection
考古研究中的系统脊椎动物收藏:田纳西大学比较脊椎动物收藏的稳定和重组
- 批准号:
9408643 - 财政年份:1994
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Reorganization and Computerization of Fossil Vertebrate Collection
脊椎动物化石收藏的重组和计算机化
- 批准号:
8611690 - 财政年份:1987
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Modernization, Reorganization and Curation of the Fossil Vertebrate Collection: Phase 2
脊椎动物化石收藏的现代化、重组和管理:第二阶段
- 批准号:
8514227 - 财政年份:1986
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
SURFACE REORGANIZATION OF THE ACTIVATED VERTEBRATE EGG
活化脊椎动物卵的表面重组
- 批准号:
3314463 - 财政年份:1984
- 资助金额:
$ 50万 - 项目类别:
SURFACE REORGANIZATION OF THE ACTIVATED VERTEBRATE EGG
活化脊椎动物卵的表面重组
- 批准号:
3314464 - 财政年份:1984
- 资助金额:
$ 50万 - 项目类别:
Modernization, Reorganization and Curation of the Fossil Vertebrate Collection: Phase I
脊椎动物化石收藏的现代化、重组和管理:第一阶段
- 批准号:
8201214 - 财政年份:1982
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Reorganization of the Vertebrate Paleontology Collection
古脊椎动物馆藏的重组
- 批准号:
8109911 - 财政年份:1981
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Reorganization of the Vertebrate Paleontological Collection of the Museum of Natural History, University of Kansas
堪萨斯大学自然历史博物馆古脊椎动物收藏的重组
- 批准号:
8115956 - 财政年份:1981
- 资助金额:
$ 50万 - 项目类别:
Continuing grant
Reorganization of the Vertebrate Paleontology Collection in The Museum of Comparative Zoology
比较动物学博物馆古脊椎动物藏品的重组
- 批准号:
7723032 - 财政年份:1978
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
Reorganization and Rehabilitation of the Vertebrate Paleontological Collections
古脊椎动物收藏的重组和恢复
- 批准号:
7680743 - 财政年份:1977
- 资助金额:
$ 50万 - 项目类别:
Continuing Grant