Structure and Inheritance of the endoplasmic reticulum

内质网的结构和遗传

• 批准号: 7216511
• 负责人: PETER Jay NOVICK
• 金额: $ 5.93万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7216511
• 关键词: actins; binding proteins; binding sites; biochemical evolution; calcium flux; cell component structure /function; cell cycle; cell line; confocal scanning microscopy; cytoplasmic receptors; endoplasmic reticulum; fluorescence recovery after photobleaching; fungal proteins; immunoelectron microscopy; microinjections; myosins; nuclear membrane; protein binding; protein localization; yeasts

DESCRIPTION (provided by applicant): In all eukarotic cells, the endoplasmic reticulum (ER) forms a highly fenestrated network that spreads throughout the volume of the cell, often lying just below the plasma membrane. We have taken a genetic approach towards understanding the mechanism of ER dynamics, structure and inheritance in yeast. Our work to date shows that ER tubules form from the nuclear envelope at the start of the cell cycle, they are delivered into the bud along actin cables by the type V myosin, Myo4p, they are anchored at the bud tip by the exocyst complex and then spread along the cell cortex to form the cortical ER. We propose that Rtn1 p, a member of the conserved reticulon family of proteins, functions as the receptor for the exocyst on the ER membrane and that Rtnlp also serves to link the ER to the cell cortex. We further propose that ergosterol and sphingolipids play an important role controlling the mobility or activity of a key component of the ER inheritance machinery at the cell cortex. Five specific aims are planned: 1) We will analyze the interaction of Rtn1 p with the exocyst, defining the topology of Rtnlp within the ER membrane and the specific protein-protein interactions that link the exocyst to Rtnlp. 2) We will determine if Rtnlp binds to a protein along the cortex and if this interaction underlies the unique localization of Rtnlp as well as the tight association of the ER with the cell cortex. 3) We will determine if Rtnlp plays a role in forming the fenestrated reticulum characteristic of the ER in all eukaryotes and if it interacts with other components of the ER membrane in this capacity. 4) We will use the information from our analysis of Rtnlp in yeast to construct dominant negative alleles of mammalian reticulon proteins. Phenotypic analysis will determine if the roles of Rtn1 p are conserved. 5) We will determine if ergosterol and sphingolipids are critical for the localization, mobility or activity of a cortical component of the machinery that establishes the structure, dynamics and inheritance of the ER.

描述（由申请人提供）：在所有真核细胞中，内质网（ER）形成一个高度有窗的网络，该网络遍布整个细胞体积，通常位于质膜正下方。我们采用遗传学方法来了解酵母内质网动态、结构和遗传的机制。迄今为止，我们的工作表明，内质网小管在细胞周期开始时从核膜形成，它们通过 V 型肌球蛋白 Myo4p 沿着肌动蛋白缆输送到芽中，它们通过外囊复合物锚定在芽尖，然后沿着细胞皮层扩散，形成皮层ER。我们认为 Rtn1 p 是保守的网状蛋白家族的成员，其功能是作为 ER 膜上的外囊的受体，并且 Rtnlp 还用于将 ER 连接到细胞皮质。我们进一步提出，麦角甾醇和鞘脂在控制细胞皮质内质网遗传机制关键成分的移动性或活性方面发挥着重要作用。计划有五个具体目标：1）我们将分析 Rtn1 p 与外囊的相互作用，定义内质网膜内 Rtnlp 的拓扑结构以及将外囊与 Rtnlp 连接的特定蛋白质-蛋白质相互作用。 2) 我们将确定 Rtnlp 是否与皮质上的蛋白质结合，以及这种相互作用是否是 Rtnlp 独特定位以及 ER 与细胞皮质紧密关联的基础。 3) 我们将确定 Rtnlp 是否在形成所有真核生物 ER 的有窗网状特征中发挥作用，以及它是否以这种能力与 ER 膜的其他成分相互作用。 4) 我们将利用对酵母中 Rtnlp 的分析信息来构建哺乳动物网织蛋白的显性失活等位基因。表型分析将确定 Rtn1 p 的作用是否保守。 5) 我们将确定麦角甾醇和鞘脂对于建立内质网结构、动态和遗传机制的皮质成分的定位、移动性或活性是否至关重要。