Isolation and identification of the gene induced by Ets family transcription factor PU.1 in murine erythroleukemia cells

小鼠红白血病细胞Ets家族转录因子PU.1诱导基因的分离与鉴定

• 批准号: 14571005
• 负责人: YAMADA Toshiyuki
• 金额: $ 1.92万
• 依托单位: Sasaki Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571005/
• 关键词: PU.1; calcium-calmodulin-dependent kinase I-like kinase (CKLiK); murine erythroleukemia(MEL); cell growth; apoptosis; PU.1; 細胞分化

PU.1, a hematopoitic cell-specific Ets family transcription factor, is involved in the generation of marine erythroleukemia (MEL). We previously reported that overexpression of PU.1 inhibits erythroid differentiation in MEL cells. To identify the target gene(s) of PU.1 in MEL cells, we carried out differential display (DD) analysis and subsequently isolated a novel gene whose expression was up-regulated in MEL cells after overexpression of PU.1. Because an 1.1 kb of open reading frame near the 5' end of the 8kb of transcript of the gene exhibited about 90% homology with the human calcium-calmodulin-dependent kinase I-like kinase (CKLiK) gene, which has been, reported to be predominantly expressed in granulocytes, it was identified as a mouse homologue of human CKLiK gene. The mouse CKLiK (mCKLiK) gene was mapped, to the mouse chromosome 2A1-A3 region, and shown to be expressed predominantly in T cells and embryonal carcinoma cells. Two types of transcripts were present showing a difference in the 3' portion of the coding region. overexpression of each isoform of mCKLiK in MEL cells revealed that one of them induces, while the other inhibits, apoptosis under low serum condition. Inhibition of erythroid-differentiation which were previously observed by us after overexpression of PU.1 in MEL cells, however, were not provoked in the cells overexpressing mCKLiK. These results suggest that mCKLiK is up-regulated by overexpression of PU.1 in MEL cells and involved in apoptosis of the cells.

PU.1是一种造血细胞特异性Ets家族转录因子，参与海洋性红白血病（MEL）的发生。我们以前报道过PU.1的过表达抑制MEL细胞中的红系分化。为了鉴定MEL细胞中PU.1的靶基因，我们进行了差异显示（DD）分析，随后分离了一种新的基因，其表达在过表达PU.1后在MEL细胞中上调。由于该基因转录本的5'端有一个1.1kb的开放阅读框，与人钙-钙调蛋白依赖性激酶I样激酶（CKLiK）基因有90%的同源性，因此被认为是人CKLiK基因在小鼠中的同源物。小鼠CKLiK（mCKLiK）基因定位于小鼠染色体2A 1-A3区域，并且显示主要在T细胞和胚胎癌细胞中表达。存在两种类型的转录物，其在编码区的3'部分中显示出差异。MEL细胞中mCKLiK的每种亚型的过表达揭示了它们中的一种在低血清条件下诱导而另一种抑制细胞凋亡。然而，我们先前在MEL细胞中过表达PU.1后观察到的红系分化抑制在过表达mCKLiK的细胞中没有引起。这些结果表明，在MEL细胞中，mCKLiK通过PU.1的过表达而上调，并参与细胞的凋亡。

项目成果

Sakurai T. et al.: "Effects of overexpression of the Ets family transcription factor TEL on cell growth and differentiation of K562 cells."Int.J.Oncol.. 22. 1327-1333 (2003)

Sakurai T.等人：“Ets家族转录因子TEL过表达对K562细胞生长和分化的影响。”Int.J.Oncol.. 22. 1327-1333 (2003)

山田俊幸: "PU.1はリンパ球系前駆細胞においてインターロイキン7受容体の発現を調節している."分子細胞治療. 2. 138-139 (2003)

Toshiyuki Yamada：“PU.1 调节淋巴祖细胞中白细胞介素 7 受体的表达。”分子细胞疗法。

Sakurai T., Yamada T et al.: "Effects of overexpression of the Ets family transcription factor TEL on cell growth and differentiation of K562 cells."Int.J.Oncol.. 22. 1327-1333 (2003)

Sakurai T.、Yamada T 等：“Ets 家族转录因子 TEL 过表达对 K562 细胞生长和分化的影响。”Int.J.Oncol.. 22. 1327-1333 (2003)

Yamada T: "PU.1 regulates expression of the interleukin-7 receptor in lymphoid progenitors. (in Japanese)"Cell.Mol.Med.. 2. 138-139 (2003)

Yamada T：“PU.1 调节淋巴祖细胞中白细胞介素 7 受体的表达。（日语）”Cell.Mol.Med.. 2. 138-139 (2003)

Suzuki M. et al.: "Direct association between PU.1 and MeCP2 that recruits mSin3A-HDAC complex for PU.1-mediated transcriptional repression."Oncogene. 22. 8688-8698 (2003)

Suzuki M. 等人：“PU.1 和 MeCP2 之间的直接关联招募 mSin3A-HDAC 复合物以进行 PU.1 介导的转录抑制。”癌基因。