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.

概括 在胚胎发育和谱系规范过程中，整合了一组复杂的细胞外信号 主转录因子 (TF) 对染色质的影响可改变染色质结构和基因表达。 PRDM 蛋白质是 TF，在其 N 末端含有 PR 甲基转移酶结构域，在其 N 末端含有 DNA 结合锌指结构域。 他们的C末端。所有 17 位 PRDM 家族成员都因其作为血统主要监管者的角色而闻名 规范及其失调的表达通常与疾病有关。 我们的小组已经表征了PRDM10（一种转录激活剂）协调小鼠的功能 植入前发育。我们的初步数据表明：1) Prdm10 合子 KO 胚胎具有可见的 胚胎日缺陷 E3.5； 2) Prdm10 母体 (Zona Pellucida3 (Zp3)-CRE) KO 胚胎在 受精卵/双细胞阶段； 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 共激活子结构域位于其 C 末端。基于这些初步数据，我们提出以下实验： 在目标 1 中，我们将确定母体 PRDM10 的功能。我们将利用 Zp3-CRE 菌株，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 的功能并确定该基因 网络直接受PRDM10监管。 在 Aim2 中，我们将确定 PRDM10 作为转录因子的作用机制。为了这个目标我们 将表征 PRDM10 的作用模式（即其甲基转移酶活性、其蛋白质相互作用组和 它的转录激活和 DNA 结合特性）。我们还将专门介绍PRDM10的功能 决定替代启动子用途的选择。最后，我们将验证卵母细胞和早期的功能 关键 PRDM10 下游效应子的胚胎发生。 这些研究的意义在于 Prdm10 是一种未表征的母体效应基因，而知识 锌指转录因子调节途径的研究将有助于干细胞生物学和生殖领域 药品。