"Conserved Cell Death Pathways in Mammals and Yeast"

描述（由申请人提供）：Bcl-2与癌症之间的关系已经被发现了20年。但是，为什么在滤泡性淋巴瘤的易位断点上发现的Bcl-2，为什么在许多肿瘤类型中Bcl-xL表达升高，以及为什么编码Bcl-2同源物的疱疹病毒导致癌症的生化机制仍然未知。该领域已经集中了大量的精力来了解抗死亡（Bcl-2和Bcl-xL）和促死亡（Bax和Bak）家族蛋白之间的复杂相互作用，以及它们在细胞受到诱导凋亡的刺激后的作用机制。我们实验室的工作迫使我们从另一个方向思考，追求Bcl-2家族蛋白的基本生化功能，这种功能在接受死亡刺激之前起作用，我们预测这种功能在抗死亡和促死亡家族成员之间是共享的。这些“日常工作”也被预测可以解释人类Bcl-2蛋白在包括哺乳动物、植物和真菌在内的多种物种中的抗死亡功能，因此，我们建议应用酵母可用的新工具来研究Bcl-2家族蛋白的“核心”功能。首先，我们寻找酵母中调控程序性细胞死亡的基因。几种不同的死亡刺激，包括病毒、热休克和代谢应激，将在区分抗和促死亡基因的条件下应用于酵母敲除菌株的完整文库。将区分特定途径的共同或独特因素。这些途径将被排序，并且已经存在的细胞死亡的候选酵母调节因子将被进一步研究，以确定它们调节线粒体形态和功能的能力之间的联系。最后，我们将对酵母进行探测，以确定人类Bcl-2和Bcl-xL抑制酵母细胞死亡所需的基因。我们预计这些新发现的酵母因子将参与线粒体的维持、生物发生和降解，并与生物能量学的调节密切相关。

DESCRIPTION (provided by applicant): The association between Bcl-2 and cancer has been known for 20 years. But the biochemical mechanisms to explain why Bcl-2, found at translocation breakpoints in follicular lymphomas, why elevated Bcl-xL expression in many tumor types, and why herpes viruses that encode Bcl-2 homologues cause cancer, remains unknown. The field has focused considerable effort to understand the complex interactions between anti-death (Bcl-2 and Bcl-xL) and pro-death (Bax and Bak) family proteins, and their mechanisms of action after cells have received a stimulus that induced apoptosis. The work in our lab has forced us to think in another direction, to pursue the basic biochemical functions of Bcl-2 family proteins that function prior to receipt of a death stimulus, and that we predict are shared between both anti- and pro-death family members. These 'day-jobs' are also predicted to explain the anti-death functions of human Bcl-2 proteins in a remarkable diversity of species including mammals, plants and fungi, Therefore, we propose to apply the new tools available for yeast to study the 'core1 functions of Bcl-2 family proteins. First, we seek to identify genes that regulate programmed cell death in yeast. Several different death stimuli, including viruses, heat shock and metabolic stress, will be applied to the complete library of yeast knockout strains under conditions that distinguish anti- and pro-death genes. Factors that are common or unique to specific pathways will be distinguished. These pathways will be ordered and candidate yeast regulators of cell death already on hand will be further investigated to determine the connection between their ability to regulate mitochondria morphology and function. Finally, yeast will be probed to identify those genes that are required for human Bcl-2 and Bcl-xL to inhibit cell death in yeast. We expect that these newly unidentified yeast factors will be involved in the maintenance, biogenesis and degradation of mitochondria, and in close proximity to the regulation of bioenergetics.