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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 急性前临床细胞白血病（APL）是由染色体15和17之间的特定相互易位引起的。易位融合了PML肿瘤抑制剂和视黄酸受体A（RARA）的基因，产生PML-RARA酶癌蛋白，与肌动杆菌的信号相关。 APL患者接受了全反式视黄酸（ATRA）的治疗，但由于突变在融合蛋白RARA部分的配体结合结构域（LBD）中抗ATRA耐药性而始终经历复发。该突变会影响蛋白质结合视黄酸的能力，并分别释放和结合核共抑制剂（N-COR）和共激活剂（ACTR）蛋白质。这些辅助因子在RARA转录函数中起着重要的作用。我们开发了荧光各向异性和共振能量转移测定法，以定量共同激活因子和核压肽与野生型RARA LBD的结合，以及在不同浓度的ATRA存在下的四个抗ATRA抗性突变体。通过使用这些测定，我们已经确定了辅助因子解离常数，并根据辅助募集来定义这些患者和细胞系突变中ATRA抗性的基础。