Novel targets of the Roberts Syndrome acetyltransferase Esco2/Eco1

罗伯茨综合征乙酰转移酶 Esco2/Eco1 的新靶标

基本信息

批准号：
10045794
负责人：
ROBERT SKIBBENS
金额：
$ 46.9万
依托单位：
LEHIGH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10045794
关键词：
Acetylation Acetyltransferase Anaphase Aneuploidy Apoptosis BACH1 gene BRCA1 gene Binding Biochemical Bioinformatics Biological Models Bone Growth Bypass Cells Chromatin Chromatin Structure Chromosome Segregation Cleft Palate Cognition Cognitive Complex Congenital Abnormality Coupled DNA DNA Damage DNA Repair DNA biosynthesis Dependence Deposition Development Embryo Etiology Failure Gastrointestinal tract structure Genes Genetic Genetic Diseases Genetic Transcription Hand Head Heart Homologous Gene Human Human Development In Vitro Intellectual functioning disability Microcephaly Mitotic Modeling Modification Mutate Mutation Nature Pathway interactions Phenotype Phocomelia Physical condensation Play Positioning Attribute Process Proxy Reaction Recombinant DNA Reporting Research Roberts-SC phocomelia syndrome Role S Phase Seizures Signal Pathway Sister Chromatid Structure Substrate Interaction Syndactyly Testing Time Tissues Translating Urinary tract Yeasts Zebrafish appendage base cohesin cohesion dosage embryo tissue enzyme substrate experience gene product genetic approach genome-wide helicase high risk human tissue in vivo innovation novel novel strategies programs reproductive tract stem cell proliferation tissue/cell culture undergraduate student yeast genetics

项目摘要

Project Summary Robert syndrome (RBS) is a severe genetic disorder characterized by phocomelia (flipper-like appendages), microcephaly, cleft palate, syndactyly, intellectual disabilities, seizures, and abnormalities in the heart and urinary, genital, and alimentary tract tissues. Importantly, mutation of ESCO2 is the single and only requirement necessary to elicit this multi- tissue and systemic suite of phenotypes. It follows that elucidating the mechanisms through which Esco2 functions will significantly advance our understanding of pathway integrations required for human development - from the physical to the cognitive. Esco2 (yeast Eco1) is an acetyltransferase that activates the DNA tethering complex - cohesin. Esco2/Eco1 and cohesin are both essential for proper chromosome segregation. In turn, the prevailing model of RBS (ESCO2 mutated) is that birth defects arise from mitotic failure (massive aneuploidy) and subsequent loss (apoptosis) of proliferative stem cells and developing tissues. Previously, we posited an alternate model in which RBS arises from transcription dysregulation. A collaborative effort indeed discovered that Esco2 and cohesin co-regulate the transcription of genes critical for bone growth. Thus, testing new and alternate models of Esco2 function will significantly advance our understanding of human development. Whether predicated on mitotic failure/apoptosis or transcriptional dysregulation, all models of RBS focus on Esco2 acetylation of the cohesin subunit Smc3. The multi-faceted nature of RBS, however, suggests that additional signaling pathways are at play. Here, we report new evidence that Eco1 (yeast homolog of Esco2) may target substrates beyond cohesins. Characterization of these novel Esco2/Eco1 substrates, and testing their roles specifically in development, provides the first novel approach to understanding RBS, and numerous other developmental maladies, to emerge in recent years. The Specific Aims of this proposal are interdependent and contain conceptually distinct genetic approaches from which to identify novel substrates of Eco1 acetylation reactions. The technical aspects of the two screens (an unbiased genome-wide cohesin-bypass screen and a genome-wide conditional dosage lethality screen) are complete – with genetic interactors in hand. In each aim, we validate the candidates, assess the acetylation state of the gene products and dependency on Eco1, and test the importance of substrate acetylation in yeast. We then translate findings into zebrafish embryos from which we can directly test for roles in development and also RBS cells to test for changes in targeted substrate modifications.

项目概要罗伯特综合征 (RBS) 是一种严重的遗传性疾病，其特征为海藻症（鳍状附属物）、小头畸形、腭裂、并指畸形、智力障碍、癫痫发作以及心脏、泌尿道、生殖道和消化道组织的异常。重要的是，ESCO2 的突变是引发这种多重突变所需的唯一条件。组织和系统表型套件由此阐明了机制。 Esco2 的哪些功能将显着增进我们对通路整合的理解人类发展所需的——从身体到认知。 Esco2（酵母 Eco1）是一种乙酰转移酶，可激活 DNA 束缚复合物 - Esco2/Eco1 和粘连蛋白对于正确的染色体分离都是必需的。反过来，RBS（ESCO2突变）的流行模型是出生缺陷是由有丝分裂失败引起的（大量非整倍性）以及随后增殖干细胞和发育干细胞的损失（凋亡）之前，我们提出了另一种模型，其中 RBS 源自转录。一项合作研究确实发现 Esco2 和粘连蛋白共同调节对骨骼生长至关重要的基因的转录因此，测试 Esco2 的新模型和替代模型。功能将极大地增进我们对人类发展的理解。无论是基于有丝分裂失败/细胞凋亡还是转录失调，所有 RBS 模型关注粘连蛋白亚基 Smc3 的 Esco2 乙酰化。然而，RBS 的性质表明还有其他信号通路在起作用。报告新证据表明 Eco1（Esco2 的酵母同源物）可能靶向超出这些新型 Esco2/Eco1 底物的表征，并测试它们的作用。特别是在开发中，提供了第一个理解 RBS 的新颖方法，并且近年来出现了许多其他发育疾病。该提案的具体目标是相互依存的，并且包含概念上不同的内容遗传方法用于识别 Eco1 乙酰化反应的新底物。两个筛选的技术方面（无偏见的全基因组粘连蛋白旁路筛选和全基因组条件剂量致死率筛查）已完成 - 具有遗传相互作用因子在每个目标中，我们都会验证候选者，评估基因产物的乙酰化状态。和对 Eco1 的依赖性，并测试酵母中底物乙酰化的重要性。将研究结果转化为斑马鱼胚胎，我们可以直接测试其中的作用开发以及 RBS 细胞来测试目标底物修饰的变化。