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）：在所有真主性细胞中，内质网（ER）形成了一个高度传针的网络，该网络在整个细胞的体积中传播，通常位于质膜下方。我们采取了一种遗传方法来理解酵母中ER动力学，结构和遗传的机制。迄今为止，我们的工作表明，在细胞周期开始时，从核神属纤维形成了ER小管，它们沿肌动蛋白电缆沿肌动蛋白型肌动蛋白（MyO4p）递送到芽中，它们通过exocyst络合物将其固定在芽尖，然后沿细胞皮质扩散，形成皮质ER。我们建议RTN1 P是蛋白质保守的网状家族的成员，它是ER膜上胞外囊肿的受体，并且RTNLP也可以将ER链接到细胞皮层。我们进一步提出，麦角固醇和鞘脂在控制细胞皮质的ER遗传机械的关键成分的迁移率或活动中起着重要作用。计划了五个具体的目的：1）我们将分析RTN1 P与胞外囊肿的相互作用，从而定义了ER膜内RTNLP的拓扑结构以及将外囊肿与RTNLP联系起来的特定蛋白质 - 蛋白质相互作用。 2）我们将确定RTNLP是否沿皮质结合，以及这种相互作用是否是RTNLP的独特定位以及ER与细胞皮质的紧密结合的基础。 3）我们将确定RTNLP是否在所有真核生物中形成ER的发否网状特征以及是否以这种能力与ER膜的其他成分相互作用。 4）我们将使用我们对酵母中RTNLP分析的信息来构建哺乳动物网状蛋白的显性负等位基因。表型分析将确定RTN1 P的作用是否保守。 5）我们将确定麦角固醇和鞘脂对于建立ER的结构，动力学和遗传的机械的皮质成分的定位，移动性或活性至关重要。