FUNCTIONAL CHARACTERIZATION OF THE PRDM10-ZN FINGER TRANSCRIPTION FACTOR IN EARLY MAMMALIAN DEVELOPMENT

PRDM10-ZN 指转录因子在早期哺乳动物发育中的功能特征

• 批准号: 10477941
• 负责人: Ernesto Guccione
• 金额: $ 36.34万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-31 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10477941
• 关键词: AT Rich Sequence; Address; Adult; Aging; Apoptosis; Automobile Driving; Binding; Biological Models; Catalytic Domain; Cell Cycle Progression; Cell physiology; Cells; Chromatin; Chromatin Structure; Communication; Complex; Couples; Cues; DNA; DNA Binding; DNA Binding Domain; Data; Defect; Developed Countries; Development; Developmental Process; Diagnosis; Disease; Drops; Embryo; Embryonic Development; Event; Family; Family Planning; Family member; Fertility; Fingers; Future; Gene Activation; Gene Expression; Genes; Genetic; Genetic Transcription; Germ Cells; Glutamine; Herpes zoster disease; Infertility; Knowledge; Link; Lysine; Malignant Fibrous Histiocytoma; Methyltransferase; Mitotic; Modeling; Mus; Nature; Oncogenic; Oocytes; Oogenesis; Pathologic; Pathway interactions; Patients; Pharmacology; Phenotype; Pre-implantation Embryo Development; Pregnancy Complications; Process; Property; Proteins; Regulation; Reproductive Biology; Reproductive Medicine; Research; Role; SET Domain; Science; Signal Transduction; Societies; Soft tissue sarcoma; Time; Tissues; Transcription Coactivator; Transcriptional Activation; Transcriptional Regulation; Translational Regulation; Translations; Vertebrates; Work; base; embryonic stem cell; epigenetic regulation; experimental study; extracellular; gene network; idiopathic infertility; inhibitor; member; paralogous gene; programs; promoter; protein expression; side effect; stem cell biology; transcription factor; zygote

SUMMARY During embryonic development and lineage specification, a complex set of extracellular signals is integrated on chromatin by Master Transcription Factors (TF) that modify chromatin structure and gene expression. PRDM proteins are TFs, containing a PR-methyltransferase domain at their N-terminus and DNA-binding Zn-fingers at their C-terminus. All 17 PRDM family members are well known for their role as master regulator of lineage specification and their deregulated expression is often linked to diseases. Our group has characterized the function of PRDM10, a transcriptional activator, orchestrating mouse preimplantation development. Our preliminary data indicate that: 1) Prdm10 zygotic KO embryos have visible defects at embryonic day E3.5; 2) Prdm10 maternal (Zona Pellucida3 (Zp3)-CRE) KO embryos arrest at the zygote/two-cell stage; 3) PRDM10 is unique among PRDM family members, as it contains a Glutamine-rich coactivator domain at its C-terminus. Based on this preliminary data we propose the following experiments: In Aim1 we will determine the function of maternal PRDM10. We will take advantage of the Zp3-CRE strain, to delete Prdm10 in the oocyte, and deplete the PRDM10 maternal protein contribution in the zygote. This will allow us to study the function of PRDM10 before zygotic transcriptional activation and determine the gene network directly regulated by PRDM10. In Aim2 we will determine the mechanism of action of PRDM10 as a transcription factor. In this aim we will characterize the mode of action of PRDM10 (i.e. its methyltransferase activity, its protein interactome, and its transcriptional activation and DNA binding-properties). We will also specifically address PRDM10’s function in dictating the choice of alternative promoter usage. Finally, we will validate the function in oocyte and early embryogenesis of key PRDM10-downstream effectors. The significance of these studies is that Prdm10 is an uncharacterized maternal effect gene, and knowledge of the pathways regulated by this Zn-finger TF will be useful to the field of stem cell biology and reproductive medicine.

摘要 在胚胎发育和谱系确定的过程中，整合了一组复杂的细胞外信号 通过主转录因子(TF)改变染色质结构和基因表达的染色质。PRDM 蛋白质是转录因子，在其N端含有PR-甲基转移酶结构域，在其N端含有与DNA结合的锌指 它们的C-末端。所有17个PRDM家族成员都是众所周知的血统主要调节者 它们的规范和表达不受控制往往与疾病有关。 我们的团队已经确定了PRDM10的功能，它是一种转录激活剂，协调小鼠 植入前发育。我们的初步数据表明：1)Prdm10合子KO胚胎具有明显的 胚胎3.5天的缺陷；2)Prdm10母体(Zp3-Cre)KO胚胎在胚胎发育早期停滞 受精卵/双细胞期；3)PRDM10在PRDM家族成员中是独一无二的，因为它含有丰富的谷氨酰胺 共激活域在其C-末端。基于这一初步数据，我们提出了以下实验： 在Aim1中，我们将确定母体PRDM10的功能。我们将利用Zp3-CRE 菌株，以删除卵母细胞中的Prdm10，并耗尽受精卵中PRDM10母体蛋白的贡献。这 将使我们能够在合子转录激活之前研究PRDM10的功能并确定基因 由PRDM10直接监管的网络。 在AIM2中，我们将确定PRDM10作为转录因子的作用机制。为了实现这一目标，我们 将表征PRDM10的作用方式(即它的甲基转移酶活性、它的蛋白质相互作用体和 其转录激活和DNA结合特性)。我们还将专门针对PRDM10的S函数 在口述替代启动子用途的选择时。最后，我们将在卵母细胞和早期验证该功能。 关键的PRDM10下游效应器的胚胎发生。 这些研究的意义在于Prdm10是一个未知的母性效应基因，并且知道 该锌指转铁蛋白调控的信号转导通路将在干细胞生物学和生殖领域发挥重要作用 医药。