Analysis of transcription factor abnormality in the progression of preleukemic state.

白血病前期状态进展中转录因子异常的分析。

• 批准号: 08671215
• 负责人: MITANI Kinuko
• 金额: $ 1.6万
• 依托单位: Tokyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671215/
• 关键词: RNA polymerase II elongation factor; elongation factor; MEN; t(11 ; 19); leukemia; AP-1 activity; 骨髄異形成症候群; t(11;19)(q.23;P13.1); MLL; 転写因子; 伸長因子

We have cloned an MIL/MEN(also termed as ELL)chimeric gene generated by the t(11 ; 19) (q23 ; p13.1), which is usually found in acute myeloid leukemia or myelodysplastic syndrome-derived leukemia. The MIL/MEN fusion protein contains the amino-terminal half of MLL including AT hook motifs which is fused to the MEN protein with a lysine-rich sequence(K.Mitani, et al. BLOOD 85 ; 2017,1995). Thus, the Mll/MEN fusion protein could be a chimeric transcription factor. Polyclonal antibodies to MLL and MEN were raised in rabbits against GST fusion of MLL(645-974 a.a.)and maltose-binding fusion of MEN(100-179 a.a.), respectively. These antibodies detected the MLL/MEN fusion protein of 230 kD,truncated form of the MLL Without the zinc finger domain and its downstream sequence(tMLL)of 180kD,and MEN protein of 70kD in Cos1 cells transfected with the corresponding cDNAs. Using immunostaining assay and subcellular fractionation of Cos1 cells expressing the MLL/MEN fusion, tMLL,and MEN proteins, we ha … More ve demonstrated that all these proteins exist in the nucleus of the cells.Conaway et al.have reported that the MEN is an RNA polymerase II elongation factor (SCIENCE 271 ; 1873,1996). The function of another elongation factor, elongin, is known to be inhibited by VHL tumor suppressor protein in vitro, suggesting the possible relationship of aberrant transcriptional elongation to oncogenesis. To demonstrate the transforming activity of the MEN protein, the MEN cDNA was introduced retrovirally into Ratl cells. Men-overexpressing cells acquired capacity of anchorage independent growth. In addition, the growth factor requirement was decreased in these cells. However, cells expressing a deletion mutant of MEN lacking the lysine-rich region did not exhibit such biological abilities. The c-Fos protein expression and AP-1 activity were elevated in the MEN-expressing cells, which might be part of the responsible mechanism for the transformation. The c-fos mRNA,the expression of which is known to be regulated partly at the stage of transcriptional elongation, appeared earlier in the MEN-expressing cells than in cells transfected with an empty vector or the deletion mutant lacking the lysine-rich region after the stimulation with epidermal growth factor. Overexpression or aberrant expression of MEN may play an important role in the leukemic tranfformation of stem cell disease. Less

我们克隆了由 t(11 ; 19) (q23 ; p13.1) 产生的 MIL/MEN（也称为 ELL）嵌合基因，该基因通常见于急性髓系白血病或骨髓增生异常综合征衍生的白血病。 MEN 融合蛋白包含 MLL 的氨基末端一半，包括 AT 钩基序，该基序与具有富含赖氨酸的 MEN 蛋白融合序列(K.Mitani, et al. BLOOD 85; 2017,1995) 因此，Mll/MEN 融合蛋白可能是一种嵌合转录因子，在兔子中产生了针对 MLL (645-) GST 融合的 MLL 和 MEN 多克隆抗体。分别为 974 个氨基酸）和 MEN 的麦芽糖结合融合体（100-179 个氨基酸）。 230 kD 的 MLL/MEN 融合蛋白、不含锌指结构域的 MLL 及其下游序列 (tMLL) 180kD 和 70kD 的 MEN 蛋白在用相应 cDNA 转染的 Cos1 细胞中使用免疫染色测定和亚细胞分级分离。 Cos1 细胞表达 MLL/MEN 融合蛋白、tMLL 和 MEN 蛋白，我们已经证明所有这些蛋白Conaway等人报道MEN是一种RNA聚合酶II延伸因子(SCIENCE 271；1873，1996)。已知另一种延伸因子elongin的功能被VHL肿瘤抑制。体外抑制蛋白，表明异常转录延伸与肿瘤发生的可能关系 为了证明 MEN 蛋白的转化活性，通过逆转录病毒引入了 MEN cDNA。此外，MEN 过表达细胞获得了贴壁独立生长的能力，但表达缺乏赖氨酸丰富区域的 MEN 缺失突变体的细胞却没有表现出这种生物学能力。 MEN 表达细胞中的 c-Fos 蛋白表达和 AP-1 活性升高，这可能是 c-fos mRNA 转化的部分机制，已知其表达在转录阶段在表皮生长因子刺激后，表达 MEN 的细胞比用空载体或缺乏富含赖氨酸区域的缺失突变体转染的细胞更早出现伸长，MEN 的过度表达或异常表达可能在白血病中发挥重要作用。干细胞疾病的转化较少。

项目成果

Antisense Repression of Proto-oncogene c-Cbl Enhances Activation of the JAK-STAT Pathway but Not the Ras Pathway in Epidermal Growth Factor Receptor Signaling*

原癌基因 c-Cbl 的反义抑制可增强表皮生长因子受体信号传导中 JAK-STAT 通路的激活，但不会增强 Ras 通路的激活*

• DOI: 10.1074/jbc.272.13.8739
• 发表时间: 1997-03-28
• 期刊: The Journal of Biological Chemistry
• 作者: H. Ueno;K. Sasaki;K. Miyagawa;H. Honda;K. Mitani;Y. Yazaki;H. Hirai
• 通讯作者: H. Hirai

The AML1/ETO(MTG8) and AML1/Evi-1 leukemia-associated chimeric oncoproteins accumulate PEBP2beta(CBFbeta) in the nucleus more efficiently than wild-type AML1.

AML1/ETO(MTG8) 和 AML1/Evi-1 白血病相关嵌合癌蛋白比野生型 AML1 更有效地在细胞核中积累 PEBP2beta(CBFbeta)。

• DOI: 10.1182/blood.v91.5.1688.1688_1688_1699
• 发表时间: 1998-09-14
• 期刊: Blood
• 影响因子: 20.3
• 作者: K. Tanaka;T. Tanaka;M. Kurokawa;Y. Imai;S. Ogawa;K. Mitani;Y. Yazaki;H. Hirai
• 通讯作者: H. Hirai