Structure and function of proteasome biogenesis associated proteins 1 and 2

描述（由申请人提供）：调节细胞内蛋白质通量是细胞代谢的关键组成部分，该过程中的缺陷与多种人类疾病有关，包括神经变性、囊性纤维化和癌症。在真核生物中，20 S蛋白酶体在细胞质和细胞核中进行大部分蛋白质水解，并且在蛋白质质量控制、信号转导、细胞周期进展、DNA修复和抗原呈递等多种过程中至关重要。双重对称真核生物20 S由28个（14 x 2）亚基组成，形成约730 kDa。桶状结构，其中包含六个蛋白水解位点。复杂的结构使得真核生物20 S不能自组装。最近，异源二聚体蛋白酶体相关生物发生因子（酵母中的Pba 1/2;哺乳动物中的PAC 1/2）已被发现作为20 S组装分子伴侣。有趣的是，Pba 1/2还包含一个C-末端HbYX序列，这是一个已建立的20 S激活基序，我们的初步数据表明，Pba 1/2结合组装的20 S蛋白酶体。我们的主要目标是确定20 S-Pba 1/2相互作用的结构基础，并朝着这个目标，我们结晶了Pba 1/2- 20 S复合物，并收集了X射线衍射数据，分辨率为3.0E。我们将使用即将到来的结构信息，以指导遗传和生化研究的功能重要性的Pba 1/2- 20 S在酵母中的相互作用。我们还将确定Pba 1/2是否与20 S组装中间体相互作用，例如1-亚基和其他分子伴侣的子集。成功的结果将提供结构和功能的洞察蛋白酶体功能一般和Pba 1/2特别。 公共卫生相关性：蛋白酶体在细胞代谢的许多方面发挥重要作用，并且是治疗干预的既定靶标，特别是用于治疗癌症。蛋白酶体功能的关键是通过结合α-亚基的末端环介导蛋白酶体的活化和调节组装的复合物。这项研究将提高对蛋白酶体结合伴侣Pba 1/2的理解，重点是结构和功能数据，这将为未来推进蛋白酶体调控领域的努力奠定基础。

DESCRIPTION (provided by applicant): Regulating intracellular protein flux is a critical component of cellular metabolism and defects in this process are linked to multiple human diseases including neurodegeneration, cystic fibrosis, and cancers. In eukaryotes, the 20S proteasome performs the bulk of proteolysis in the cytosol and nucleus and is vital in diverse processes such as protein quality control, signal transduction, cell cycle progression, DNA repair, and antigen presentation. The two-fold symmetric eukaryotic 20S is composed of 28 (14 x 2) subunits that form a ~730 kDa. barrel-like structure, which contains the six proteolytic sites. The complicated architecture renders eukaryotic 20S incapable of self-assembly. Recently, a heterodimeric Proteasome Associated Biogenesis factor (Pba1/2 in yeast; PAC1/2 in mammals) has been discovered that acts as a 20S assembly chaperone. Interestingly, Pba1/2 also contains a C-terminal HbYX sequence, which is an established 20S activating motif, and our preliminary data show that Pba1/2 binds to the assembled 20S proteasome. Our primary objective is to determine the structural basis for the 20S-Pba1/2 interaction, and toward this goal we have crystallized a Pba1/2-20S complex and collected X-ray diffraction data to 3.0E resolution. We will use the forthcoming structural information to guide genetic and biochemical investigations into the functional importance of Pba1/2-20S interactions in yeast. We will also determine if Pba1/2 interacts with 20S assembly intermediates, such as subsets of 1-subunits and other chaperones. Successful outcome will provide structural and functional insight into proteasome function in general and Pba1/2 in particular. PUBLIC HEALTH RELEVANCE: Proteasomes play essential roles in many facets of cellular metabolism and are established targets for therapeutic intervention, especially for the treatment of cancers. Critical to proteasome function are complexes that mediate the activation and regulated assembly of proteasomes by binding their end rings of alpha-subunits. This study will improve understanding of the proteasome-binding partner Pba1/2, with an emphasis on structural and functional data that will lay the foundation for future efforts to advance the field of proteasome regulation.