INDUCTION AND BIOGENESIS OF SUBCELLULAR ORGANELLES

亚细胞器的诱导和生物发生

• 批准号: 3268808
• 负责人: SAMUEL KAPLAN
• 金额: $ 3.68万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-01 至 1990-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3268808
• 关键词: Rhodopseudomonas; bacterial genetics; bacterial proteins; cell membrane; electron microscopy; enzyme mechanism; gel electrophoresis; genetic manipulation; immunochemistry; intracellular membranes; lipid biosynthesis; lipid structure; membrane lipids; membrane proteins; membrane reconstitution /synthesis; membrane structure; nucleic acid probes; organelles; phospholipids; photosynthetic bacteria; protein structure; protein transport

Employing the facultative photoheterotrophic bacterium, Rhodopseudomonas sphaeroides, we hope to understand those genetic, physiologic and biochemical events which are involved in the localized intracellular differentiation, induced by anaerobiosis, of the cytoplasmic membrane resulting in the formation of a physically contiguous but structurally and functionally new membrane system (ICM). We further hope to understand how the cell maintains this differentiated state by an in vitro analysis of those events which direct newly synthesized proteins to either the cytoplasmic membrane, the ICM or to both. Similarly, we hope to understand the maintenance of the structural and functional integrity of the ICM by investigating: 1. the role of phospholipid exchange proteins and phospholipid biosynthetic activities in the movement of phospholipids from their site of synthesis to the ICM at precisely pre-determined times in the cellcycle, 2. the role of lipid structure and/or protein structure in the maintenance of the structural and functional isolation of the ICM, 3. the functional dependency of the ICM on the structural relationships of its interacting components through the use of specific cross-linking reagents, immunological, electron microscopic analyses and detailed biochemical and physiological studies of isolated functional components from wild type and mutant organisms. Finally, we shall investigate using immunochemical, molecular genetic, biochemical and physiological techniques the fate of the ICM and its components when growing cells are returned to conditions which no longer favor the maintenance of the ICM. The application of specific genetic probes will facilitate these studies by permitting us to investigate the transcriptional regulation of ICM development and maintenance effected by O2 as well as adjustments in the intracellular levels and composition of the ICM effected by light intensity. A thorough understanding of membrane structure, function, and synthesis is essential to an understanding of the secretory, energetic, conductance and selective permeability properties of membranes observed in normal and diseased tissue.

利用兼性光异养细菌红假单胞菌 sphaeroides，我们希望了解这些遗传，生理和 参与局部细胞内的生化事件 细胞质膜的分化，由厌氧诱导 导致形成物理上连续但结构上和 功能性新膜系统（ICM）。 我们还希望了解 细胞通过体外分析保持这种分化状态， 这些事件将新合成的蛋白质引导到 细胞质膜，ICM或两者。 同样，我们希望了解 通过以下方式维持ICM的结构和功能完整性 调查：1.磷脂交换蛋白的作用 磷脂生物合成活动中的磷脂从 它们的合成位点在ICM中精确预定的时间， cellcycle，2.脂质结构和/或蛋白质结构在 保持ICM的结构和功能隔离，3.的 ICM的功能依赖于其结构关系， 通过使用特定的交联剂， 免疫学、电子显微镜分析和详细的生物化学和 对来自野生型的分离的功能性组分的生理学研究， 变异生物 最后，我们将使用免疫化学， 分子遗传学、生物化学和生理学技术， 当生长细胞返回到 不再支持ICM的维护。 实施具体 基因探针将通过允许我们 研究ICM发育的转录调控， 维持受O2影响以及调节细胞内 ICM的水平和组成受光照强度的影响。 的透彻 了解膜的结构、功能和合成是必不可少的 对分泌、能量、传导和选择性的理解 在正常和患病组织中观察到的膜的渗透性特性。