SCF Ubiquitin Ligases in Cell Cycle Control and Chromosome Stability

SCF 泛素连接在细胞周期控制和染色体稳定性中的作用

• 批准号: 10599187
• 负责人: Michael James Emanuele
• 金额: $ 31.44万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10599187
• 关键词: Address; Amyotrophic Lateral Sclerosis; Apoptosis; Attention; Binding; CRISPR/Cas technology; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Line; Cells; Cementation; Chromosomal Stability; Clinical; Complex; Cyclin-Dependent Kinase Inhibitor; Cyclin-Dependent Kinases; Cyclins; Data; Deubiquitination; Disease; Enzymes; Eukaryota; F Box Domain; F-Box Proteins; Family; Funding; Gene Expression; Genes; Genetic Transcription; Goals; Growth; Health; Human; Human Cell Line; Impairment; Kinetics; Knowledge; Ligase; Link; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; Methodology; Mus; Mutation; Names; Neurodegenerative Disorders; Normal Cell; PLK1 gene; Pathway interactions; Phenotype; Phosphoproteins; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Post-Translational Regulation; Process; Proliferating; Protein Family; Proteins; Proteome; Proteomics; RBL2 gene; Regulation; Resources; Retinoblastoma; Role; S phase; System; Textbooks; Tumor Suppressor Proteins; Ubiquitin; Ubiquitin family; Ubiquitination; Work; cdc Genes; cyclin F; genome integrity; genome-wide; human disease; in silico; innovation; loss of function; member; nervous system disorder; ovarian neoplasm; overexpression; programs; protein degradation; receptor; retinoblastoma pathway; senescence; ubiquitin isopeptidase; ubiquitin ligase; ubiquitin-protein ligase

PROJECT SUMMARY ABSTRACT Dynamic oscillations in the abundance and activity of key proteins drives cell cycle progression. This is typified by cyclins and cyclin kinase inhibitors (CKIs), which oscillate during cell cycle progression and determine the activation kinetics of Cyclin Dependent Kinases, which propel the cell cycle forward. Defining the pathways, networks and mechanisms underlying dynamics in protein abundance during normal cell cycles is essential to understanding proliferative control. Ubiquitin is the major regulator of protein degradation in eukaryotes and plays an essential and highly conserved role in cell cycle progression. The Skp1-Cul1-F-box (SCF) family of E3 ubiquitin ligases are major regulators of cell cycle progression and sculpt the protein landscape post- translationally to facilitate proliferation. SCF ligases engage a set of ~70 interchangeable substrate receptors termed F-box proteins, which dictate SCF target selection. The eponymous Cyclin F is the founding member of the F-box family and has a cyclin homology domain like that found in the canonical CDK activators. Rather than activate a CDK to promote cell cycle via phosphorylation, cyclin F binds to the SCF, promoting cell cycle through ubiquitination. Cyclin F mRNA and protein levels oscillate significantly during the cell cycle and to a greater extent than other F-box encoding genes. Cyclin F is required for viability in mice and essential for growth/survival in many human cell lines. Further, it is overexpressed in cancer, and its mutation is linked to the neurodegenerative disease amyotrophic lateral sclerosis. Nevertheless, there remain significant knowledge gaps related to what pathways cyclin F controls and how it is regulated. Defining substrates and mechanisms of cyclin F is significant to a fundamental understanding of cell cycle, as well as human health. During the previous funding cycle, our lab identified several cyclin F substrates, highlighting its importance in cell cycle control. Here, we build on that work, defining a new substrate with important roles in normal cell cycle control and with relevance to disease, while also beginning to dissect the complex mechanisms by which cyclin F is controlled. Borne out of innovative, unbiased, computational, and proteomic strategies, our data suggest new, critical roles for cyclin F in cell cycle, and undescribed mechanisms underlying its regulation. In aim 1, we examine the role of cyclin F in G1/S progression through regulation of a tumor suppressor in the retinoblastoma (RB)-pocket protein family, RBL2/p130. We will determine the mechanisms by which cyclin F controls the RB-pathway and how this contributes to gene expression, cell cycle, and proliferative control. In aims 2 and 3, we turn our attention towards mechanisms that converge on cyclin F to regulate its abundance and potentially activity. Aim 2 is focused on the regulation of cyclin F by kinases and how they determine cyclin F stability. In aim 3 we extend this analysis of post-translational regulation of cyclin F, determining how it is regulated by deubiquitinases. Together, these studies will provide a comprehensive analysis for how cyclin F is itself regulated and its contributions to cell cycle in humans.

项目摘要摘要 关键蛋白的丰度和活性中的动态振荡驱动细胞周期进程。这是指的 通过细胞周期蛋白和细胞周期蛋白激酶抑制剂（CKI），它们在细胞周期进程中振荡并确定 细胞周期蛋白依赖性激酶的激活动力学，这些激酶会推动细胞周期前进。定义路径， 正常细胞周期期间蛋白质丰度动态的网络和机制对于 了解增生性控制。泛素是真核生物和蛋白质降解的主要调节剂 在细胞周期进程中起着必不可少的和高度保守的作用。 E3的SKP1-CUL1-F-BOX（SCF）家族 泛素连接酶是细胞周期进程的主要调节剂，并在 翻译以促进增殖。 SCF连接酶接合一组约70个可互换的底物受体 称为F-box蛋白，该蛋白决定SCF目标选择。同名细胞周期蛋白F是 F-box家族具有类似于规范CDK激活剂中的细胞周期蛋白同源性域。相当 比激活CDK以通过磷酸化促进细胞周期，Cyclin F与SCF结合，促进细胞周期 通过泛素化。细胞周期蛋白F mRNA和蛋白质水平在细胞周期中显着振荡，并且 比其他F-box编码基因更大。细胞周期蛋白F是在小鼠中的生存力所必需的，对于 在许多人类细胞系中的生长/生存。此外，它在癌症中过表达，其突变与 神经退行性疾病肌萎缩性侧硬化症。然而，仍然有重要的知识 与Cyclin F控制途径及其调节方式有关的差距。定义底物和机制 细胞周期蛋白F对细胞周期以及人类健康的基本了解至关重要。在 以前的资金周期，我们的实验室确定了几个细胞周期蛋白F底物，突出了其在细胞周期中的重要性 控制。在这里，我们以这项工作为基础，定义了在正常细胞周期控制中具有重要作用的新底物 并且与疾病有关，同时也开始剖析细胞周期蛋白F的复杂机制 受控。我们的数据是由创新，公正，计算和蛋白质组学策略所证实的，我们的数据表明了新的 细胞周期蛋白F在细胞周期中的关键作用，以及其调节的未描述机制。在AIM 1中，我们 通过调节肿瘤抑制因子在G1/S进展中的作用 视网膜母细胞瘤（RB） - 口袋蛋白家族，RBL2/P130。我们将确定细胞周期蛋白F的机制 控制RB-Pathway以及这如何促进基因表达，细胞周期和增殖对照。在 目标2和3，我们将注意力转向融合细胞周期蛋白F以调节其丰度的机制 和潜在的活动。 AIM 2的重点是通过激酶调节细胞周期蛋白F及其如何确定细胞周期蛋白 F稳定性。在AIM 3中，我们扩展了对细胞周期蛋白F的翻译后调节的分析，确定了它的状态 由去泛素酶调节。这些研究将共同​​为细胞周期蛋白F的方式提供全面的分析 本身受到调节及其对人类细胞周期的贡献。