MOLECULAR BASIS OF ACUTE PROMYELOCYTIC LEUKEMIA

急性早幼粒细胞白血病的分子基础

• 批准号: 6377993
• 负责人: J DON CHEN
• 金额: $ 31.59万
• 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/6377993
• 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：(改编自调查人员的摘要)t(15；17) 易位占所有人类急性早幼粒细胞白血病的95%以上 白血病(APL)病例。这种易位将PML基因与核融合在一起 维甲酸受体(RARA)，并产生致癌融合蛋白， PML-RARA。目前认为PML-RARA可能干扰了PML-RARA的功能。 野生型PML和RARA。免疫细胞化学研究揭示了细胞的定位 PML位于离散的核结构域，称为PML致癌结构域(Pod)，Kr 遗体、ND10或核体。当吊舱的确切功能保持不变 不清楚，它的结构在t(15；17)易位的APL细胞中被破坏， 这表明豆荚在APL中发挥了重要作用。要研究其功能 在豆荚中，他们已经使用酵母筛选出PML相互作用的因子 双杂交系统。他们已经分离出三种特定的相互作用蛋白 包括先前证明PML修饰剂SUMO-1和两种新的 参与转录调控的蛋白质。在这个项目中， 他们将研究PML与这两个新辅助因子的相互作用。 标准技术包括酵母双杂交试验，GST下拉， Far-Western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。基于他们的 初步研究表明，这两个辅因子是转录因子。 抑制者。将进行实验以确定其作用机制。 这些蛋白质的转录抑制作用。他们提出，人类的角色 PML和Pod将这些辅因子招募到受限的PML区域 通过与PML的蛋白质-蛋白质相互作用。他们进一步建议本地化 这些辅因子对豆荚的作用将导致它们的失活 转录抑制因子功能。这些假设将通过一项 瞬时转基因和免疫细胞化学研究相结合。在……里面 此外，他们将调查这些PML辅助因子在APL中的作用，基于 观察到这两种蛋白也与致癌的APL相互作用 融合蛋白，PML-RARA。总之，这些研究应该会给我们带来新的曙光 PML的功能及其核域，以及提供新的见解 新辅因子的功能和调节。更好地理解 APL在分子和细胞水平的机制应导致更多的 本病的有效诊断和治疗，以及提供 对其他恶性肿瘤的影响。