Histone proline isomerization and gene regulation

组蛋白脯氨酸异构化和基因调控

• 批准号: 7566297
• 负责人: Nobuaki Kikyo
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-20 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7566297
• 关键词: Acetylation; Affect; Amino Acids; Autologous Transplantation; Binding; Biochemical; Biological; Biological Models; Cell Differentiation process; Cell Nucleus; Cells; Chromatin; Cloning; Complement; Complex; Cytoplasm; DNA; DNA Microarray Chip; Data; Disease; Egg Proteins; Employee Strikes; Epigenetic Process; FOS gene; Fibroblasts; Fluorescence; Fluorescence Recovery After Photobleaching; Future; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Histone H3; Histones; In Vitro; Incubated; Isomerase; JUN gene; Lead; Life; Lysine; Malignant Neoplasms; Mediating; Methods; Methylation; Mitogens; Modification; Molecular; Molecular Chaperones; Movement; Mus; Nuclear; Paper; Patients; Peptide Conformation; Peptide Fragments; Peptidylprolyl Isomerase; Phosphorylation; Photobleaching; Physiological; Play; Preparation; Process; Proline; Proteins; Publishing; Reaction; Recording of previous events; Regenerative Medicine; Relaxation; Reporting; Repression; Research; Research Personnel; Role; Series; Somatic Cell; Stem cells; Tail; Techniques; Technology; Testing; Tissues; Transplantation; Xenopus; Xenopus oocyte; base; cell dedifferentiation; chromatin protein; chromatin remodeling; egg; embryonic stem cell; histone modification; induced pluripotent stem cell; innovation; insight; interest; novel; nuclear reprogramming; nucleoplasmin; pluripotency; public health relevance; stem cell biology; tool; transcription factor

DESCRIPTION (provided by applicant): Existence of powerful nuclear reprogramming activities in egg cytoplasm has been unequivocally demonstrated by successful somatic cell nuclear cloning using terminally differentiated cell nuclei as donors. These reprogramming activities can induce dedifferentiation of the donor nuclei, leading to their acquisition of a topipotent state. Despite the long history of the study and the particularly intensive recent research, however, currently little is known about the molecular mechanism behind nuclear reprogramming during cloning. The long-term goal of this project is to identify these egg-derived nuclear reprogramming factors and apply the factors to reprogram the differentiation status of living cells. This project potentially leads to efficient preparation of patients-derived new tissues for autologous transplantation, complementing the recently reported iPS cell technology. Global chromatin decondensation is one of the most striking cell biological changes observed in the donor nuclei. The investigator's group has recently found that the egg protein nucleoplasmin and the peptidyl isomerase PPIA are responsible for the chromatin decondensation. These two proteins trigger acetylation, phosphorylation and methylation on specific histones and promote gene activation. Based on these observations, it was hypothesized that nucleoplasmin and PPIA organize a specific set of epigenetic modifications on the target genes, which leads to chromatin relaxation and nuclear reprogramming. To test this hypothesis, following specific aims are proposed. In Specific Aim 1 the relationship among phosphorylation, methylation and isomerization of histones will be investigated by using peptide fragments, chromatin and living cells. In Specific Aim 2, how nucleoplasmin and PPIA are involved in nuclear reprogramming and pluripotency in embryonic stem cells will be studied by genetic knockdown, DNA microarray and the fluorescence photobleaching technique. In addition, promotion of cell dedifferentiation by these factors will be investigated in combination with stem cell-specific transcription factors. Together, these studies will provide a novel insight into the reprogramming of cell differentiation through chromatin remodeling. PUBLIC HEALTH RELEVANCE: These studies will substantially contribute to our understanding on the regulation of gene activity, which is important for the therapy of many diseases including cancer. In addition, the results will facilitate the progress of nuclear reprogramming and regenerative medicine, especially creation of patients-derived rejection-free tissues for the purpose of transplantation.

描述（由申请人提供）： 通过终止分化的细胞核作为供体，成功证明了卵细胞质中强大的核重编程活性的存在。这些重编程活性可以诱导供体核的去分化，从而导致它们获得托管状态。尽管这项研究的历史悠久，并且最近进行了特别密集的研究，但目前对克隆过程中核重编程背后的分子机制知之甚少。该项目的长期目标是确定这些鸡蛋衍生的核重编程因素，并应用这些因素来重新编程活细胞的分化状态。该项目有可能导致有效制备患者衍生的新组织进行自体移植，并补充了最近报道的IPS细胞技术。全局染色质脱凝性是供体核中观察到的最引人注目的细胞生物学变化之一。研究者组最近发现，卵子蛋白核纤维蛋白和肽基异构酶PPIA负责染色质脱敏。这两种蛋白质会触发乙酰化，磷酸化和甲基化，并促进基因激活。基于这些观察结果，假设核纤维蛋白和PPIA在靶基因上组织了一组特定的表观遗传修饰，这会导致染色质弛豫和核重编程。为了检验该假设，提出了以下特定目标。在特定的目标1中，将使用肽片段，染色质和活细胞来研究组蛋白的磷酸化，组蛋白的甲基化和异构化之间的关系。在特定的目标2中，将通过遗传敲低，DNA微阵列和荧光光漂白技术研究核纤维素和PPIA如何参与胚胎干细胞中的核重编程和多能性。此外，将研究这些因素对细胞去分化的促进，将与干细胞特异性转录因子结合使用。总之，这些研究将通过染色质重塑对细胞分化的重编程提供新的见解。公共卫生相关性：这些研究将有助于我们对基因活性调节的理解，这对于包括癌症在内的许多疾病的治疗非常重要。此外，结果将促进核重编程和再生医学的进展，尤其是为了移植而创建患者衍生的无排斥组织。