MOLECULAR BASIS OF ACUTE PROMYELOCYTIC LEUKEMIA

急性早幼粒细胞白血病的分子基础

• 批准号: 6802828
• 负责人: J DON CHEN
• 金额: $ 31.49万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6802828
• 关键词: DNA binding protein; acute myelogenous leukemia; apoptosis; cell differentiation; chimeric proteins; chromosome translocation; gene induction /repression; genetic transcription; immunofluorescence technique; immunoprecipitation; intermolecular interaction; molecular oncology; neoplasm /cancer genetics; oncoproteins; retinoid binding proteins; tissue /cell culture; transcription factor; transport proteins; yeast two hybrid system

DESCRIPTION: (Adapted from the investigator's abstract) The t(15;17) translocation accounts for over 95 percent of all human acute promyelocytic leukemia (APL) cases. This translocation fuses the PML gene with the nuclear receptor for retinoic acid (RARa) and creates an oncogenic fusion protein, PML-RARa. It is thought that PML-RARa may interfere with the function of wild-type PML and RARa. Immunocytochemistry studies reveal the localization of PML at discrete nuclear domains known as PML oncogenic domains (PODs), Kr bodies, ND10, or nuclear bodies. While the exact function of the PODs remains unclear, its structure is disrupted in t(15;17)-translocation APL cells, suggesting an important roles for the PODs in APL. To investigate the function of the PODs, they have screened for PML-interacting factors using the yeast two-hybrid system. They have isolated three specific interacting proteins including a previously demonstrated PML-modifier, SUMO-1, and two novel proteins that are implicated in transcriptional regulation. In this project, they will investigate the interactions of PML with these two novel cofactors. Standard techniques including the yeast two-hybrid assay, GST pull down, Far-Western blot, and immunofluorescence microscopy will be utilized to establish the interaction of PML with these cofactors. Based on their preliminary studies, they propose that these two cofactors are transcriptional repressors. Experiments will be conducted to determine the mechanism of transcriptional repression by these proteins. They propose that the roles of PML and the PODs are to recruit these cofactors to the restricted PML domains by protein-protein interaction with PML. They further propose that localization of these cofactors to the PODs will lead to inactivation of their transcriptional repressor function. These hypotheses will be tested by a combination of transient transfection and immunocytochemistry studies. In addition, they will investigate the role of these PML cofactors in APL, based on observations that these two proteins also interact with the APL oncogenic fusion protein, PML-RARa. Together, these studies should shed new lights into the function of PML and its nuclear domains, as well as providing new insights to the function and regulation of the new cofactors. A better understanding of mechanism of APL at the molecular and cellular levels should lead to a more effective diagnosis and treatment of this disease, as well as providing implications for other malignancies.

描述：（根据研究者的摘要进行了改编）t（15; 17） 易位占所有人类急性临时细胞的95％以上 白血病（APL）病例。这种易位将PML基因与核基因融合 视黄酸的受体（RARA），并创建致癌融合蛋白， PML拉拉。人们认为PML-RARA可能会干扰 野生型PML和RARA。免疫细胞化学研究揭示了 在离散的核域下的PML称为PML致癌域（POD），KR 身体，ND10或核体。虽然豆荚的确切功能仍然存在 尚不清楚，其结构在t（15; 17） - 换位apl细胞中被破坏， 提出了apl中豆荚的重要作用。调查功能 在豆荚中，它们已经使用酵母进行了筛选的PML相互作用因子 两个杂交系统。他们已经隔离了三种特定的相互作用蛋白 包括先前证明的PML模型，SUMO-1和两个小说 与转录调控有关的蛋白质。在这个项目中， 他们将研究PML与这两个新型辅助因子的相互作用。 标准技术，包括酵母双杂交测定法，GST下拉， 遥远的印迹和免疫荧光显微镜将用于 建立PML与这些辅助因子的相互作用。基于他们的 初步研究，他们提出这两个辅助因子是转录的 阻遏物。将进行实验以确定 这些蛋白质的转录抑制。他们建议 PML和POD将招募这些辅助因子到受限的PML域 通过蛋白质 - 蛋白质与PML相互作用。他们进一步提出了本地化 这些辅助因子到豆荚中将导致其失活 转录阻遏函数。这些假设将通过 瞬时转染和免疫细胞化学研究的组合。在 此外，他们将研究这些PML辅助因子在APL中的作用，基于APL 关于这两种蛋白质也与APL致癌性相互作用的观察 融合蛋白，PML-RARA。这些研究在一起应该把新的灯弄掉 PML及其核领域的功能以及提供新的见解 对新辅助因子的功能和调节。更好地理解 APL在分子和细胞水平上的机制应导致更多 有效诊断和治疗该疾病，并提供 对其他恶性肿瘤的影响。