Deubiquitinases in Cell Cycle Control

细胞周期控制中的去泛素酶

• 批准号: 10725061
• 负责人: Michael James Emanuele
• 金额: $ 3.93万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10725061
• 关键词: Address; Aging; Binding; Biochemical; Biochemistry; Cardiac health; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Proliferation; Cell division; Cell physiology; Cells; Cellular biology; Chromosomal Stability; Chromosome Segregation; Data; Data Set; Disease; Drug Targeting; Enzymatic Biochemistry; Enzymes; Family; Future; G1/S Transition; Genome; Genome Stability; Health; Human; Infection; Kinetics; Knowledge; Ligase; Maintenance; Malignant Neoplasms; Mitotic; Mitotic Checkpoint; Molecular Biology; Normal Cell; Outcome; Pathologic; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Phenotype; Play; Polyubiquitin; Positioning Attribute; Proliferating; Proteins; Proteome; Proteomics; Regulation; Role; S phase; Signal Transduction; Specific qualifier value; Substrate Interaction; System; Techniques; Therapeutic; Ubiquitin; Ubiquitination; Work; cancer cell; genome integrity; in silico; inhibitor; loss of function; multicatalytic endopeptidase complex; nervous system disorder; ovarian neoplasm; protein degradation; restraint; therapeutic target; ubiquitin isopeptidase; ubiquitin-protein ligase; virtual

DEUBIQUITINASES IN CELL CYCLE CONTROL Project Summary Ubiquitin signaling contributes to virtually all aspects of cell physiology and is implicated in aging and disease. The covalent conjugation of polyubiquitin chains onto substrates triggers their degradation by the proteasome, as well as various other cellular outcomes. Ubiquitination is carried out by an enzymatic cascade of ubiquitin activators (E1), conjugators (E2) and ligases (E3). During normal cell cycles the ubiquitin system plays an essential and conserved role in remodeling the protein landscape. Ubiquitin substrates are determined by E3 ligases and much of our understanding of ubiquitin signaling has focused on the identity, substrates and mechanisms of E3s. However, ubiquitination is reversible, and ubiquitin is removed from substrates by catalytic proteases termed DUBs (deubiquitinases). Despite their critical role in sculpting the proteome, much less is known about the identity, substrates and mechanisms of DUBs in cell cycle progression, when compared to their E3 counterparts. Nevertheless, dysregulation of both E3s and DUBs alters cell cycle progression and has deleterious effects on genome integrity. Moreover, both E3s and DUBs can be perturbed in pathologies such as cancer, contributing to the biochemical and phenotypic features of disease. Thus, defining the identity, substrates and mechanisms of DUBs in cell cycle is essential to understanding how normal cell proliferation and genome stability are maintained and coordinated. We hypothesize that DUBs are essential for cell cycle progression and chromosome stability and are equally important as their E3 counterparts. We address this hypothesis in three specific aims, that combine complementary techniques, and which focus on the role of DUBs in major cell cycle transitions. In Aims 1 and 2 we investigate Cezanne/OTUD7B, an ovarian tumor family deubiquitinase, that we recently demonstrated is cell cycle regulated and which controls the M to G1 transition. In Aim 1, we will determine substrates for Cezanne using proteomics approaches, define mechanisms of DUB-substrate interactions, and the role of Cezanne in the degradation of substrates at M/G1. In Aim 2, we will expand this analysis to determine how Cezanne is itself regulated, both at the level of its abundance and activity, and then determine how these regulatory systems influence its role in cell division. Finally, in Aim 3, we determine the role of DUBs in a second major cell cycle transition, G1/S. The G1/S boundary is a major barrier to proliferation in normal and cancer cell cycles and relies heavily on ubiuqitin signaling. However, little is known about DUBs involved in G1/S. We will use computational approaches and loss-of-function screens, to identify and then investigate DUBs that control G1/S. Collectively, this proposal will fill significant knowledge gaps in the cell cycle, ubiquitin and DUB fields, related to roles and mechanisms of DUBs in proliferation and genome maintenance. My lab is uniquely positioned to address these questions, illustrated by our prior work, that includes global analysis of ubiquitination networks, detailed analysis of specific ubiquitin pathways and their role in cell cycle, and determination of the function and enzymology of cell cycle DUBs.

细胞周期控制中的去泛素酶 项目摘要 泛素信号传导几乎有助于细胞生理的所有方面，并与衰老和疾病有关。 多泛素链在底物上的共价结合触发了蛋白酶体的降解， 以及其他各种细胞结局。泛素化是通过泛素的酶促级联 激活剂（E1），偶联器（E2）和连接酶（E3）。在正常细胞周期中，泛素系统播放 在重塑蛋白质景观中的重要和保守的作用。泛素底物由E3确定 连接酶和我们对泛素信号传导的大部分理解都集中在身份，底物和底物上 E3的机制。但是，泛素化是可逆的，泛素通过催化从底物中除去 蛋白酶称为配音（去泛素酶）。尽管它们在雕刻蛋白质组中的作用至关重要，但更少的是 与其相比 E3对应物。然而，E3S和DUB的失调都会改变细胞周期的进展，并具有 对基因组完整性的有害影响。此外，E3和配音在诸如病理中都可能受到干扰 癌症，有助于疾病的生化和表型特征。因此，定义身份，底物 细胞周期中配音的机制对于了解正常细胞增殖和基因组是至关重要的 保持稳定性并协调。我们假设配音对于细胞周期进程至关重要 染色体稳定性，与E3对应物同样重要。我们在三个中解决了这一假设 具体目的，结合了互补技术，并关注配音在主要细胞周期中的作用 过渡。在目标1和2中，我们调查了卵巢肿瘤家族去泛素酶Cezanne/otud7b，我们 最近证明的是细胞周期调节并控制M到G1转变。在AIM 1中，我们将 使用蛋白质组学方法来确定塞尚的底物，定义配音底物的机制 相互作用，以及塞尚在m/g1下底物降解中的作用。在AIM 2中，我们将扩展此事 分析以确定Cezanne本身如何在其丰度和活动的水平上进行调节，然后 确定这些调节系统如何影响其在细胞分裂中的作用。最后，在AIM 3中，我们确定 DUB在第二个主要细胞周期过渡中的作用G1/s。 G1/S边界是扩散的主要障碍 在正常和癌细胞周期中，严重依赖乌比素信号传导。但是，关于配音知之甚少 参与G1/s。我们将使用计算方法和功能丧失屏幕来识别然后 研究控制G1/s的配音。总的来说，该建议将填补细胞周期中的显着知识差距， 泛素和配音领域，与配音在增殖和基因组维持中的作用和机制有关。 我的实验室在我们先前的工作中说明了这些问题，包括全球 分析泛素化网络，特定泛素途径的详细分析及其在细胞周期中的作用， 并确定细胞周期配音功能和酶学。