Study of cell type specific gene expression of runt related murine genes

矮小相关小鼠基因的细胞类型特异性基因表达研究

• 批准号: 04833015
• 负责人: MARUYAMA Mitsuo
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04833015/
• 关键词: transcription factor; t(8, 21)translocation; cell type specific gene expression; AML1; 分子生物学; 遺伝子発現調節機構; 核内転写因子; 急性骨髄性白血病; DNA・結合・二量体形成ドメイン; 染色体転座

PEBP2alpha subunit is a member of a novel transcription factor family with the Runt-Domain which is responsible for its DNA binding and hetrodimerizing with beta subunit activities. Three murine PEBP2alpha genes, alphaA, alphaB and alphaC, have been identified. alphaA is originally isolated as a Polyoma virus enhancer binding protein and involved the T cell specific gene expression or early development. alphaB is the murine homologue of human AML1 which was found at the breakpoint of the t(8,21). The only gene encoding beta subunit was identified and the expression of its mRNA is ubiquitous among the cell lines we tested. The expressions of alphaA mRNAs indicate the cell type specific manner and suggest to vary its function as a transcription factor. In this study, we analyzed the expression pattern of alphaA subspecies (alphaA1 , alphaA2, alphaA3) mRNAs to characterize the machinery of cell type specificity of mRNA expression. A primary Northern blot analysis suggested that alphaA3 mRNA was minor species like alphaA2 mRNA.To confirm their expression, RT-PCR was performed and was found that the alphaA3 mRNA, like alphaA1, was expressed both in Ha-ras transformed NIH3T3 cells and murine T cell line, EL4. Moreover, little expression of alphaA3, like alphaA1, was detected in murine embryonic carcinoma cell line, P19. After the treatment of retinoic acid, however, significant induction of alphaA3 mRNA was obsered in the P19 cells. On the other hand, alphaA2 mRNA was expressed in all cell lines we tested as well as beta subunit mRNAs. These study strongly suggested that the various isoforms derived from a single gene by alternative splicing were differently regulated the expression of each mRNA.It is a valuable question we should discuss further that the biological significance of this expression specificity and the mechanism of their transcription regulation.

Pebp2alpha亚基是带有Runt域的新型转录因子家族的成员，该家族负责其DNA结合和通过β亚基活性进行Hetrodimerigation。已经鉴定出了三个鼠PEBP2ALPHA基因，Alphaa，Alphab和Alphac。 αA最初被分离为多瘤病毒增强子结合蛋白，并涉及T细胞特异性基因表达或早期发育。 Alphab是人类AML1的鼠同源物，在t的断点（8,21）。鉴定出编码β亚基的唯一基因，并且其mRNA的表达在我们测试的细胞系中无处不在。 αAmRNA的表达表明特定细胞类型的方式，并建议将其作为转录因子的功能改变。在这项研究中，我们分析了αA亚种的表达模式（Alphaa1，alphaA2，alphaA3）mRNA，以表征mRNA表达的细胞类型特异性的机械。主要的北印迹分析表明，αA3mRNA是像alphaa2 mRNA这样的次要物种。为了确认其表达，进行了RT-PCR，发现alphaA3 mRNA（如alphaa1）在HA-RAS中都表达了NIH3T3细胞和Murine T细胞系，EL4在HA-RAS中均表达。此外，在鼠类胚胎癌细胞系中检测到αA3的几乎没有表达，例如αA1。然而，在治疗视黄酸后，在p19细胞中观察到了αA3mRNA的显着诱导。另一方面，在我们测试的所有细胞系以及β亚基mRNA中表达αA2mRNA。这些研究强烈表明，通过替代剪接从单个基因得出的各种同工型对每个mRNA的表达进行了不同的调节。这是一个有价值的问题，我们应该进一步讨论，这种表达特异性的生物学意义及其转录调节的机制。

项目成果

Bae, S.-C., Ogawa, E., Maruyama, M., Oka, M., Satake, M., Shigesada, K., Jenkins, N.A., Gilbert, D.J., Copeland, N.G.and Ito, Y.: "PEBP2alphaB/mouse AML1 consists of multiple isoforms that possess differential transactivation potentials." Mol.Cell.Biol.(i

Bae, S.-C.、Okawa, E.、Maruyama, M.、Oka, M.、Satake, M.、Shigesada, K.、Jenkins, N.A.、Gilbert, D.J.、Copeland, N.G. 和 Ito, Y.：

Suk Chul Bae: "Isolation of PEBP2αB cDNA representing the Mouse homologue of human acute myeloid leukemia gene.AML1" Oncogene. 8. 809-814 (1993)

Suk Chul Bae：“代表人急性髓性白血病基因.AML1 的小鼠同源物的 PEBP2αB cDNA 的分离”Oncogene 8. 809-814 (1993)。

Eiko Ogawa: "Molecular cloning and characterization of PEBP2β.the heterodimeric partner of a novel Drosophila runt-related DNA binding protein PEBP2α" Virology. 194. 314-331 (1993)

Eiko Okawa：“PEBP2β 的分子克隆和表征。新型果蝇矮小相关 DNA 结合蛋白 PEBP2α 的异二聚体伙伴”病毒学 194. 314-331 (1993)。

Ogawa, E., Maruyama, M., Kagoshima, H., Inuzuka, M., Lu, J., Satake, M., Shigesada, K.and Ito, Y.: "PEBP2/PEA2 represents a family of transcription factors -homologous to the products of the Drosophila runt gene and the human AML1 gene." Proc.Natl.Acad Sc

Okawa, E.、Maruyama, M.、Kagoshima, H.、Inuzuka, M.、Lu, J.、Satake, M.、Shigesada, K. 和 Ito, Y.：“PEBP2/PEA2 代表转录因子家族

Eiko Ogawa: "PEBP2/PEA2 represents a new family of transcription factor homologous to the products of the Drosophila runt and human AML1" Proc.Natl.Acad.Sci.USA. 90. 6859-6863 (1993)

Eiko Okawa：“PEBP2/PEA2 代表了一个新的转录因子家族，与果蝇侏儒和人类 AML1 的产物同源”Proc.Natl.Acad.Sci.USA。