FUNCTIONAL ANALYSIS OF MLL AND MLL PARTNER FUSIONS

描述：（改编自研究者摘要）总体 本研究的目的是研究正常MLL的功能 蛋白质和由染色体易位形成的融合蛋白质 导致白血病的MLL 他们将使用几种方法， 鉴定正常MLL和两种MLL融合体的下游靶基因， MLL-AF 9和MLL-CBP。 AF 9是急性MLL中最常见的融合伴侣， 骨髓性白血病，包括新发和继发于药物治疗， 靶向DNA拓扑异构酶II。 CBP是MLL的融合合作伙伴，迄今为止， 仅在治疗相关的白血病中观察到，但有一个例外。 他们 将根据MLL的能力鉴定正常MLL的靶基因， 蛋白质与靶DNA序列或结构结合， 免疫沉淀。 从这些实验中，他们将确定 它直接或间接地与MLL结合。 他们还将定义 控制结合的参数，包括DNA靶 序列/结构，相互作用蛋白质，涉及的MLL结构域，和蛋白质 改性 此外，他们还将鉴定出表达与 通过表达MLL-AF 9或MLL-CBP融合蛋白改变， 瞬时或诱导型稳定表达系统，随后进行基因表达 使用cDNA微阵列在全基因组范围内进行模式分析， 寡核苷酸微阵列芯片 这将确定目标， 表达在白血病发生中可能是重要的，无论机制如何。 直接和下游靶点将有助于识别细胞通路 这些蛋白质被融合蛋白解除调节，并且可能对细胞的生长至关重要。 终极白血病表型 总之，这些实验中提出的 两个具体目标将提供有关该机制的宝贵信息 正常和白血病细胞中MLL和MLL-伴侣基因功能。

DESCRIPTION: (adapted from the investigator's abstract) The overall objective of this proposal is to study the function of the normal MLL protein and of the fusion proteins formed by chromosomal translocations involving MLL that result in leukemia. They will use several approaches to identify downstream target genes of normal MLL and of two MLL fusions, MLL-AF9 and MLL-CBP. AF9 is the most common fusion partner of MLL in acute myeloid leukemia, both de novo and secondary to therapy with drugs that target DNA topoisomerase II. CBP is a fusion partner of MLL that has so far been observed only in therapy-related leukemias with one exception. They will identify target genes of normal MLL based on the ability of the MLL protein to bind to a target DNA sequence or structure using immunoprecipitation. From these experiments, they will identify targets of MLL to which it binds either directly or indirectly. They will also define the parameters that control binding, which may include DNA target sequence/structure, interacting proteins, MLL domains involved, and protein modification. Additionally, they will identify genes whose expression is altered by expression of either MLL-AF9 or MLL-CBP fusion proteins using transient or inducible stable expression systems followed by gene expression pattern analysis on a genome-wide scale using cDNA microarrays and oligonucleotide microarray chips. This will identify targets whose altered expression may be important in leukemogenesis, regardless of the mechanism. The direct and the downstream targets will help identify cellular pathways that are deregulated by the fusion proteins and that may be critical for the ultimate leukemia phenotype. Together, the experiments proposed in these two specific aims will provide valuable information regarding the mechanism of MLL and MLL-partner gene function in normal and in leukemic cells.