Functional Characterization of LIMK2 carrying LIM and PDZ Domains in the Developmental and Tumorigenic Process.

携带 LIM 和 PDZ 结构域的 LIMK2 在发育和致瘤过程中的功能表征。

• 批准号: 09670170
• 负责人: OSADA Hirotaka
• 金额: $ 2.05万
• 依托单位: Aichi Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670170/
• 关键词: Lung Cancer; Oncogene; がん遺伝子

We previously isolated the human LIMK2 gene and identified two alternative transcripts, LIMK2a and LIMK2b, which were differentially regulated in a tissue-specific manner. To examine this differential tissue-specific expression in detail, an RNase protection assay was performed, which demonstrated three expression patterns of the LIMK2 isoforms. In digestive organs, the LIMK2a transcripts were preferentially expressed in fetal and adult tissues ; in brain and lung, the LIMK2a transcript was predominantly expressed only in fetal tissue, and in placenta, the LIMK2b transcript was expressed more abundantly than that of LJMK2a. To further investigate this mechanism and the transcription factors involved, we isolated the two distinct 5' upstream regions from the phage genomic library, and found that both LIMK2a and 2b promoters have a single major transcription initiation site and thecharacteristics of a TATA-less promoter. A luciferase reporter assay of the transcriptional activity revealed positive as well as negative regulatory regions within both promoters. The RORalphal may be involved in the positive regulation for the LIMK2b expression, while the MZF-1 might regulate both LIMK2a and 2b in a different manner. The genomic structure of the LIMK2 gene was also determined. These findings should lead to a better understanding of the possibly diverse functions of the LIMK family

我们之前分离了人类LIMK2基因，并鉴定了两个替代转录本，LIMK2a和LIMK2b，它们以组织特异性的方式进行差异调节。为了详细检查这种差异组织特异性表达，进行了RNase保护实验，证明了LIMK2亚型的三种表达模式。在消化器官中，LIMK2a转录本优先在胎儿和成人组织中表达；在脑和肺中，LIMK2a转录本仅在胎儿组织中主要表达，而在胎盘中，LIMK2b转录本的表达量高于LJMK2a。为了进一步研究这一机制及其涉及的转录因子，我们从噬菌体基因组文库中分离出两个不同的5'上游区域，发现LIMK2a和2b启动子都有一个主要的转录起始位点，并且具有无tata启动子的特征。荧光素酶的转录活性报告分析显示，两个启动子内都有阳性和阴性的调控区域。roral可能参与了LIMK2b表达的正向调控，而MZF-1可能以不同的方式调控LIMK2a和2b的表达。LIMK2基因的基因组结构也被确定。这些发现将有助于更好地理解LIMK家族可能具有的多种功能

项目成果

Yatabe, Y., et al.: "p27KIP1 in human lung cancers : differential changes in small cell and non-small cell carcinomas." Cancer Res.58. 1042-7 (1998)

Yatabe, Y., et al.：“人类肺癌中的 p27KIP1：小细胞癌和非小细胞癌的差异变化。”

Osada, H., et al.: "LIM-only protein Lmo2 forms a protein complex with erythroid transcription factor GATA-1" Leukemia. 11 Suppl 3. 307-12 (1997)

Osada, H. 等人：“仅 LIM 蛋白 Lmo2 与红系转录因子 GATA-1 形成蛋白复合物”白血病。

Kondo M, Osada H, et al.: "Molecular cloning of human TAK1 and its mutational analysis in human lung cancer" International Journal of Cancer. 75. 559-563 (1998)

Kondo M、Osada H 等人：“人类 TAK1 的分子克隆及其在人类肺癌中的突变分析”国际癌症杂志。

Takagi Y, Osada H, et al.: "p53 mutations in non-small cell lung cancers occurring in individuals without a past history of active smoking." British Journal of Cancer. (印刷中). (1998)

Takagi Y、Osada H 等人：“无主动吸烟史的个体中发生的 p53 突变”（正在出版）。

Osada, H.et al.: "The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNA-binding complex which includes the TAL1, E47, GATA-1 and Ldb1/NLI proteins." EMBO Journal. 16. 3142-57 (1997)

Osada, H.等人：“仅 LIM 蛋白 Lmo2 是一种桥接分子，可组装红系 DNA 结合复合物，其中包括 TAL1、E47、GATA-1 和 Ldb1/NLI 蛋白。”