MEMBRANE PROLIFERATIONS INDUCED BY HMGCOA REDUCTASE IN YEAST

酵母中 HMGCOA 还原酶诱导的膜增殖

• 批准号: 7358038
• 负责人: Randolph Y. Hampton
• 金额: $ 0.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358038
• 关键词: MEMBRANE; PROLIFERATIONS; INDUCED; HMGCOA; REDUCTASE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The endoplasmic reticulum (ER) displays extreme plasticity in a broad range of eukaryotes: the structure of the ER can vary widely in response to natural stimuli or experimental manipulations. Despite the ubiquitousness of this cellular response, little is known about the structural features or molecular underpinnings of ER plasticity. High expression levels of certain ER resident proteins cause profound proliferation of the ER membrane. The best characterized of these proteins is HMG-CoA Reductase (HMGR), which we are utilizing to rigorously examine ER plasticity in Saccharomyces cerevisiae. HMGR levels naturally fluctuate in response to the mevalonate pathway, can be increased or decreased using drugs or regulated through molecular biological mechanisms, making this an ideal system to investigate organellar dynamics. A great deal of information fundamental to understanding this biological process is attainable only by advanced light and electron microscopy techniques. For example, we can utilize electron tomography to determine if the membrane structures caused by elevated levels of HGMR are a proliferation or a structural reorganization of the ER. We can also analyze the dynamics of the system by inducing proliferations or stimulating the degradation of proliferations. Budding sites of membrane proliferations as well as structural intermediates in forming or disassembling the membrane arrays may be identified in this manner. The dynamic behavior of HMGR is completely unknown. We are able employ FRET to study the dynamics of a functional HMGR-GFP and its ability to move in and out of proliferations. Coupling the powerful genetics and biochemistry of this yeast system with advanced microscopy techniques will provide a strong platform for an enhanced understanding ER plasticity and dynamics.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。内质网（ER）在广泛的真核生物中显示出极强的可塑性：ER的结构可以在自然刺激或实验操作中发生很大变化。尽管这种细胞反应无处不在，但对ER可塑性的结构特征或分子基础知之甚少。某些ER驻留蛋白的高表达水平引起ER膜的显著增殖。这些蛋白质中最具特征的是HMG-CoA还原酶（HMGR），我们正在利用它来严格检查酿酒酵母中的ER可塑性。HMGR水平响应于甲羟戊酸途径而自然波动，可以使用药物增加或减少或通过分子生物学机制调节，使其成为研究细胞器动力学的理想系统。只有通过先进的光学和电子显微镜技术才能获得理解这一生物过程的大量基本信息。例如，我们可以利用电子断层扫描来确定由HGMR水平升高引起的膜结构是ER的增殖还是结构重组。我们还可以通过诱导增殖或刺激增殖的退化来分析系统的动力学。膜增殖的出芽位点以及形成或分解膜阵列的结构中间体可以以这种方式鉴定。HMGR的动态行为是完全未知的。我们能够利用FRET研究功能性HMGR-GFP的动力学及其移入和移出增殖的能力。将这种酵母系统的强大遗传学和生物化学与先进的显微镜技术相结合，将为增强对ER可塑性和动力学的理解提供强大的平台。