Thirman M J, Levitan D A, Kobayashi H, Simon M C, Rowley J D
Department of Medicine, University of Chicago Medical Center, IL 60637.
Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12110-4. doi: 10.1073/pnas.91.25.12110.
To characterize the functions of MLL fusion transcripts, we cloned the gene that fuses to MLL in the translocation t(11;19)(q23;p13.1). This translocation is distinct from another type of 11;19 translocation with a 19p13.3 breakpoint that results in the fusion of MLL to the ENL gene. By PCR screening of a cDNA library prepared from a patient's leukemia cells with this translocation, we obtained a fusion transcript containing exon 7 of MLL and sequence of an unknown gene. The sequence of this gene was amplified and used as a probe to screen a fetal brain cDNA library. On Northern blot analysis, this cDNA detected a 4.4-kb transcript that was abundant in peripheral blood leukocytes, skeletal muscle, placenta, and testis and expressed at lower levels in spleen, thymus, heart, brain, lung, kidney, liver, and ovary. In addition, a 2.8-kb transcript was present in peripheral blood, testis, and placenta. On "zoo blots," this gene was shown to be evolutionarily conserved in 10 mammalian species as well as in chicken, frog, and fish. We have named this gene ELL (for eleven-nineteen lysine-rich leukemia gene). A highly basic, lysine-rich motif of the predicted ELL protein is homologous to similar regions of several proteins, including the DNA-binding domain of poly(ADP-ribose) polymerase. The characterization of the normal functions of ELL as well as its altered function when fused to MLL will be critical to further our understanding of the mechanisms of leukemogenesis.
为了表征MLL融合转录本的功能,我们克隆了在t(11;19)(q23;p13.1)易位中与MLL融合的基因。这种易位不同于另一种11;19易位,后者的断点在19p13.3,导致MLL与ENL基因融合。通过用这种易位的患者白血病细胞制备的cDNA文库进行PCR筛选,我们获得了一个包含MLL外显子7和一个未知基因序列的融合转录本。扩增该基因的序列并用作探针筛选胎儿脑cDNA文库。在Northern印迹分析中,该cDNA检测到一个4.4kb的转录本,其在外周血白细胞、骨骼肌、胎盘和睾丸中丰富,在脾、胸腺、心脏、脑、肺、肾、肝和卵巢中表达水平较低。此外,在外周血、睾丸和胎盘中存在一个2.8kb的转录本。在“动物杂交印迹”中,该基因在10种哺乳动物以及鸡、青蛙和鱼中显示出进化保守性。我们将该基因命名为ELL(十一 - 十九富含赖氨酸白血病基因)。预测的ELL蛋白的一个高度碱性、富含赖氨酸的基序与几种蛋白质的类似区域同源,包括聚(ADP - 核糖)聚合酶的DNA结合结构域。表征ELL的正常功能以及其与MLL融合时功能的改变对于进一步理解白血病发生机制至关重要。