CHARACTERIZATION OF A HOMEOBOX-RELATED GENE,ATBF1-ITS POTENTIAL ROLE IN CELL GROWTH AND NEURONAL CELL DIFFERENTLATION AND ANALYSIS OF POLYMORPHISMS IN THE TRIPLET REPEAT REGIONS

同源盒相关基因ATBF1的表征及其在细胞生长和神经细胞分化中的潜在作用以及三联体重复区的多态性分析

• 批准号: 06454611
• 负责人: SHIMA Tomoko
• 金额: $ 2.82万
• 依托单位: HYOGO COLLEGE OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454611/
• 关键词: ATBF1; mouse ATBF1; alternative splicing; neuronal differentiation; granurocytic differentiation; homeodomain; tumorigenesity; triplet repeat; co-transfection; ATBF1遺伝子産物(蛋白); polyclonal抗体

I.The ATBF1 gene was originally isolated from a cDNA library derived from human hepatocellular carcinoma cells (Jpn.J.Electrophoresis). Since the alternative splicing mechanisms during cell differentiation and malignant transformaiton were found in a variety of tumor cell lines (J.Mol.Endocrinol.), we tried to isolate the ATBF1 alternative-spliced isoforms. We found a longer ATBF1-cDNA,ATBF1-A,from a cDNA library derived from human fibroblasts. The cDNA of ATBF1-A is 12kb, encoding a 400kDa protein, while the original ATBF1, now termed ATBF1-B,is 8kb encoding a 300kDa protein. ATBF1-A has 4 homeobox and 23 zinc finger domains and one of the largest gene of the homeodomain-zinc finger gene family, and its expression was highly induced during neuronal differentiation (JBC). We also cloned the mouse ATBF1-A,whose expression is high in the developing brain but low in the adult tissues (Gene). On the other hand, the expression of ATBF1-B was specifically induced during granurocytic differen … More tiation of leukemia cells. Taken together, two isoforms of ATBF1 are independently expressed in a cell specific manner, however, further investigation is needed using other cell lineage.Using rabbit polyclonal antibodies, a 250kDa protein was detected by Western bloting that the ATBF1 protein may be cleaved after transcription. Also, a possibility of dimerization of the ATBF1 at the homeodomains were found.II.The expression vectors of ATBF1 were transfected to NIH-3T3 and PC12 cells. The exogenous ATBF1 was detected just after the transfeciton, but lost after several pasages, indicating that its overexpression reduced cell growth whle non-transfectants could survive. These results suggest that the ATBF1 expression may suppress tumor growth through both growth arrest and cellular differentiation.III.There are several triplet repeat regions in the ATBF1 gene, and 6-10 repeat polymorphism was found at a CAA region. DNAs isolated from patients with several neurodegenerative diseases showed no apparent expansion of this region. The ATBF1 was mapped on chromosome 16q22, however, no linkage has been reported in this region so far. When some diseases will be mapped in this region, more study must be conducted. Less

I. ATBF1基因最初是从源自人肝细胞癌细胞（JPN.J.Electropolosisis）的cDNA文库中分离出来的。由于在各种肿瘤细胞系中发现了细胞分化和恶性转化过程中的替代剪接机制（J.Mol.endocrinol。），因此我们试图隔离ATBF1替代替代的同工型。我们从源自人成纤维细胞的cDNA库中发现了更长的ATBF1-CDNA ATBF1-A。 ATBF1-A的cDNA为12KB，编码了400KDA蛋白，而原始的ATBF1（现在称为ATBF1-B）是编码300KDA蛋白的8KB。 ATBF1-A具有4个同源词和23个锌指结构域，并且是同源区锌指基因家族的最大基因之一，其表达在神经元分化（JBC）期间被高度诱导。我们还克隆了小鼠ATBF1-A，其表达在发育中的大脑中很高，但在成年组织中（基因）中很低。另一方面，在颗粒细胞差异期间特别诱导了ATBF1-B的表达……白血病细胞更多。综上所述，ATBF1的两个同工型以细胞特异性方式独立表达，但是，使用其他细胞谱系需要进一步研究。使用兔多克隆抗体，通过蛋白质印迹检测到250kDa蛋白，即转录后可以清除ATBF1蛋白。同样，发现了同源词组中ATBF1维度的可能性。II。将ATBF1的表达向量转换为NIH-3T3和PC12细胞。在经狂热者之后检测到外源ATBF1，但在几条路径后丢失，表明其过表达降低了细胞生长，非转移剂可以生存。这些结果表明，ATBF1表达可以通过生长停滞和细胞分化来抑制肿瘤的生长。III.III.ATBF1基因中有几个三重型重复区域，在CAA区域发现了6-10个重复多态性。从几种神经退行性疾病患者中分离出的DNA显示该区域没有明显的扩张。 ATBF1在16q22染色体上绘制了映射，但是到目前为止，该地区尚未报告任何联系。当将某些疾病映射到该地区时，必须进行更多的研究。较少的

项目成果

Hashimoto,T.,et al.: "Induction of the expression of the ATBF1 gene during differentiation of hematopoetic cells" in preparation.

Hashimoto,T.,et al.：“造血细胞分化过程中 ATBF1 基因表达的诱导”正在准备中。

Miura,Y.,et al.: "Cloning and characterization of an ATBF1 isoform that expresses in a neuronal differentiation-dependent manner" J. Biol. Chem. 270. 26840-26848 (1995)

Miura,Y.,et al.：“以神经元分化依赖性方式表达的 ATBF1 同工型的克隆和表征”J. Biol。

Kodama,A.,et al.: "A new CAA repeat polymorphism in the ATBF1 gene" in preparation.

Kodama,A.,et al.：“ATBF1 基因中的新 CAA 重复多态性”正在准备中。

Hashimoto, T., et al.: "Induction of the expression of the ATBF1 gene during differentiation of leukemic cell lines." (in preparation).

Hashimoto, T. 等人：“在白血病细胞系分化过程中诱导 ATBF1 基因的表达。”

Nakamura,M.,et al.: "Multiple alternative splice isoforms of parathyroid hormone-related peptide mRNA in human cell lines" J. Mol. Endocrin.15. 245-249 (1995)

Nakamura,M.,et al.：“人类细胞系中甲状旁腺激素相关肽 mRNA 的多种选择性剪接亚型”J. Mol。