Deubiquitinases in Cell Cycle Control

细胞周期控制中的去泛素酶

• 批准号: 10559372
• 负责人: Michael James Emanuele
• 金额: $ 7.86万
• 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/10559372
• 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; Proteins; Proteome; Proteomics; Regulation; Role; S phase; Signal Transduction; Specific qualifier value; Substrate Interaction; System; Techniques; Therapeutic; Ubiquitin; Ubiquitination; Work; base; cancer cell; genome integrity; in silico; inhibitor; loss of function; multicatalytic endopeptidase complex; nervous system disorder; ovarian neoplasm; protein degradation; therapeutic target; 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.

细胞周期调控中的脱泛素酶 项目摘要 泛素信号对细胞生理学的各个方面都有影响，并与衰老和疾病有关。 多泛素链在底物上的共价结合引发了蛋白酶体对它们的降解， 以及其他各种细胞结果。泛素化是通过泛素级联酶来实现的 激活子(E1)、接合子(E2)和连接酶(E3)。在正常的细胞周期中，泛素系统发挥着 在重塑蛋白质格局中起着至关重要的和保守的作用。泛素底物由E3决定 连接酶和我们对泛素信号的大部分理解都集中在身份、底物和 E3S的作用机制。然而，泛素化是可逆的，泛素是通过催化从底物中去除的 被称为DUBS(脱泛素酶)的蛋白酶。尽管它们在塑造蛋白质组中起着关键作用，但更少的是 已知DUBS在细胞周期进程中的身份、底物和机制，当与它们的 E3对应方。然而，E3和DUBS的失调改变了细胞周期进程，并 对基因组完整性的有害影响。此外，E3和DUB都可以在病理上受到干扰，例如 癌症，导致疾病的生化和表型特征。因此，定义身份、底物 而DUBS在细胞周期中的作用机制对于理解正常的细胞增殖和基因组 保持和协调稳定。我们假设DUBS对细胞周期进程和 染色体的稳定性和它们的E3对应物同样重要。我们从三个方面阐述了这一假说 特定的目标，结合互补的技术，并专注于DUB在主要细胞周期中的作用 过渡。在目标1和2中，我们研究了Cezanne/OTUD7B，一种卵巢肿瘤去泛素酶家族，我们 最近证明是细胞周期调节的，它控制着M向G1的转变。在目标1中，我们将 用蛋白质组学方法确定塞尚的底物，确定复制底物的作用机制 相互作用，以及塞尚在M/G1底物降解中的作用。在目标2中，我们将扩展这一点 分析以确定Cezanne本身是如何受到监管的，无论是在其丰度和活动水平上，然后 确定这些调控系统如何影响其在细胞分裂中的作用。最后，在目标3中，我们确定 DUBS在第二个重要的细胞周期转换G1/S中的作用G1/S边界是细胞增殖的主要障碍 在正常和癌细胞周期中，并且严重依赖泛素信号转导。然而，人们对dubs知之甚少。 参与了G1/S。我们将使用计算方法和功能丧失筛查，以确定和然后 研究控制G1/S的DUB，总的来说，这一建议将填补细胞周期中显著的知识空白， 泛素和DUB域，与DUBS在增殖和基因组维持中的作用和机制有关。 我的实验室处于独特的位置来解决这些问题，我们之前的工作说明了这一点，其中包括全球 分析泛素化网络，详细分析特定的泛素途径及其在细胞周期中的作用， 以及细胞周期DUBS的功能和酶学的测定。