MOLECULAR BASIS OF ACUTE PROMYELOCYTIC LEUKEMIA

急性早幼粒细胞白血病的分子基础

• 批准号: 6166043
• 负责人: J DON CHEN
• 金额: $ 33.84万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 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 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6166043
• 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：（改编自研究者摘要）t（15;17） 在所有人类急性早幼粒细胞白血病中， 白血病（APL）病例。这种易位融合了PML基因与核 视黄酸受体（RARa）并产生致癌融合蛋白， PML-RARA a.据认为PML-RAR α可能会干扰 野生型PML和RAR α。免疫细胞化学研究揭示了 在称为PML致癌结构域（POD）的离散核结构域处的PML，Kr ND 10或核小体。虽然POD的确切功能仍然存在 不清楚，其结构在t（15;17）易位APL细胞中被破坏， 提示POD在APL中起重要作用。为了研究 在POD中，他们使用酵母筛选PML相互作用因子， 双混合动力系统他们已经分离出三种特定的相互作用的蛋白质 包括之前证明的PML修饰剂SUMO-1和两种新型修饰剂 参与转录调控的蛋白质。在本项目中， 他们将研究PML与这两种新型辅因子的相互作用。 标准技术包括酵母双杂交测定、GST下拉、 远蛋白质印迹和免疫荧光显微镜将用于 建立PML与这些辅因子的相互作用。基于其 初步研究表明，这两个辅因子是转录的， 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将这些辅因子募集到受限的PML结构域中 通过蛋白质-蛋白质相互作用与PML结合。他们进一步提出， 这些辅助因子的POD将导致其失活， 转录抑制因子功能。这些假设将由一个 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辅因子在APL中的作用， 观察到这两种蛋白质也与APL致癌基因相互作用， 融合蛋白PML-RAR α。总之，这些研究应该为我们提供新的视角， PML及其核结构域的功能，并提供新的见解 新辅因子的功能和调节。更好地了解 APL在分子和细胞水平上的机制应该导致更多的 有效诊断和治疗这种疾病，以及提供 对其他恶性肿瘤的影响。