THE PML-RAR ONCOGENIC FUSION PROTEIN AND ITS ROLE IN ACUTE PROMYELOCYTIC LEUKEMI

PML-RAR 致癌融合蛋白及其在急性早幼粒细胞白血病中的作用

• 批准号: 7610048
• 负责人: CHRISTOPHER S FRANCKLYN
• 金额: $ 1.32万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610048
• 关键词: Acute; Acute Promyelocytic Leukemia; Affect; Binding; Biological Assay; Cell Line; Chimeric Proteins; Chromosomes, Human, Pair 15; Computer Retrieval of Information on Scientific Projects Database; Dissociation; Energy Transfer; Fluorescence Anisotropy; Funding; Genetic Transcription; Grant; Institution; Ligand Binding Domain; Mutation; Myelogenous; Nuclear; Oncogene Proteins; Oncogenic; PML gene; Patients; Peptides; Play; Proteins; Reciprocal Translocation; Relapse; Research; Research Personnel; Resistance; Resources; Retinoic Acid Binding; Retinoic Acid Receptor; Role; Signal Transduction; Source; Tretinoin; Tumor Suppressor Proteins; United States National Institutes of Health; base; cofactor; mutant

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Acute Promyelocytic Leukemia (APL) results from a specific reciprocal translocation between chromosomes 15 and 17. The translocation fuses the genes for the PML tumor suppressor and retinoic acid receptor a (RARa), generating a PML-RARa chimeric oncoprotein that interferes with signaling associated with myeloid differentiation. APL patients receive treatment with all-trans retinoic acid (ATRA), but invariably undergo relapse owing to mutations imparting ATRA resistance in the ligand binding domain (LBD) of the RARa portion of the fusion protein. The mutations affect the ability of the protein to bind retinoic acid, as well as undergo release and binding of nuclear co-repressor (N-CoR) and co-activator (ACTR) proteins, respectively. These cofactors play essential roles in RARa transcription function. We developed fluorescence anisotropy and resonance energy transfer assays to quantitate the binding of coactivator and corepressor peptides to the wild-type RARa LBD and four ATRA resistant mutants in the presence of varying concentrations of ATRA. By use of these assays, we have determined cofactor dissociation constants and defined the basis of ATRA resistance in these patient and cell line mutations in terms of cofactor recruitment.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 急性早幼粒细胞白血病 (APL) 是由 15 号和 17 号染色体之间的特定相互易位引起的。这种易位融合了 PML 肿瘤抑制基因和视黄酸受体 a (RARa) 的基因，产生了一种 PML-RARa 嵌合癌蛋白，该蛋白干扰与骨髓分化。 APL 患者接受全反式视黄酸 (ATRA) 治疗，但由于融合蛋白 RARa 部分的配体结合域 (LBD) 中产生 ATRA 抗性突变，因此总是会出现复发。这些突变影响蛋白质结合视黄酸的能力，并分别影响核辅阻遏蛋白（N-CoR）和辅激活蛋白（ACTR）的释放和结合。这些辅助因子在 RARa 转录功能中发挥重要作用。我们开发了荧光各向异性和共振能量转移测定法，以定量在不同浓度的 ATRA 存在下，共激活剂和辅阻遏物肽与野生型 RARa LBD 和四种 ATRA 抗性突变体的结合。通过使用这些测定，我们确定了辅因子解离常数，并定义了这些患者中 ATRA 耐药性的基础以及辅因子募集方面的细胞系突变。