ANALYSES OF THE TRANSCRIPTIONAL REGULATORY NETWORK FOR DROSOPHILA DNA REPLICATION GENES

果蝇DNA复制基因转录调控网络分析

• 批准号: 11480217
• 负责人: YAMAGUCHI Masamitsu
• 金额: $ 6.02万
• 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 (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11480217/
• 关键词: DROSOPHILA; PCNA; RFX; DNA replication-related genes; Chromatin remodling factor; Mi-2; DREF; DNA復製関連遺伝子; DNA複製遺伝子; 転写制御; 転写因子

We have identified a novel transcriptional regulatory element, PRE (positive regulatory element) located between URE (upstream regulatory element) and DRE (DNA replication-related element) in the Drosophila PCNA (proliferating cell nuclear antigen) gene. One-hybrid screening using the PRE sequence as a bait allowed us to clone a CDNA encoding a novel transcription factor carrying the RFX domain. The factor was named as Drfx2 (Drosophila RFX2). Over expression of DNA-binding domain of this factor induced the abnormal eye morphology (rough eye phenotype) hi adult flies. Since the human RFX protein is reported to be involved in regulation of the human PCNA gene, function of RFX family protein may be conserved between mammal and Drosophila.We have also identified Mi-2, a chromatin remodeling factor as an interactant of the transcription factor DREF that coordinately regulates DNA replication related genes in Drosophila. Over-expression of DREF in eye imaginal discs induces ectopic DNA synthesis and apoptosis in eye disc cells and consequently the adult flies exhibit the rough eye phenotype. Half reduction of the Mi-2 gene dose strongly enhanced the DREFinduced rough eye phenotype, suggesting that Mi-2 negatively regulates the function of DREF in vivo. In consistent with this, the purified GST-Mi-2 fusion protein inhibits DNA-binding activity of DREF in vitro.

我们已经确定了一个新的转录调节元件，即果蝇PCNA（增殖细胞核抗原）基因的URE（上游调节元件）和DRE（DNA复制相关元件）之间的PRE（正调节元件）。使用前序列作为诱饵的一杂交筛选使我们能够克隆cDNA，该cDNA编码了带有RFX结构域的新型转录因子。该因子称为DRFX2（果蝇RFX2）。该因子的DNA结合结构域的表达诱导了异常的眼睛形态（粗眼表型）HI成年苍蝇。由于据报道人RFX蛋白参与了人PCNA基因的调节，因此RFX家族蛋白的功能可以在哺乳动物和果蝇之间保留。我们还确定了MI-2，这是MI-2，这是一种染色质重塑因子，是转录因子的相互作用因素，可用于协同调节DNA复制基因drosophophila中的DNA复制基因。眼睛想象盘中排水的过表达可诱导眼盘细胞的异位DNA合成和凋亡，因此成年苍蝇表现出粗糙的眼睛表型。 MI-2基因剂量的一半降低强烈增强了干燥的粗眼表型，这表明MI-2对DREF在体内的功能进行负调节。与此一致，纯化的GST-MI-2融合蛋白在体外抑制了DREF的DNA结合活性。

项目成果

Kwon,E.-J., et al.: "Transcriptional regulation of the Drosophila raf proto-oncogene by D-STAT durimg development and immune response."J.Biol.Chem.. 275. 19824-19830 (2000)

Kwon,E.-J., et al.：“D-STAT 在发育和免疫反应过程中对果蝇 raf 原癌基因的转录调节。”J.Biol.Chem.. 275. 19824-19830 (2000)

Kwon.E.-J., et al.: "Transcriptional regulation of the Drosophila raf proto-oncogene by the transcription factor dE2F."Nucleic Acids Res.. (in press). (2001)

Kwon.E.-J. 等人：“转录因子 dE2F 对果蝇 raf 原癌基因的转录调节”。《核酸研究》（正在出版）。

Ohno, K., Takahashi, Y., Hirose, F., Inoue, Y.H., Taguchi, O., Nishida, Y., Matsukage, A. and Yamaguchi, M.: "Characterization of a Drosophila homologue of the human myelodysplasia/myeloid leukemia factor (MLF)"Gene. 260. 133-143 (2000)

Ohno, K.、Takahashi, Y.、Hirose, F.、Inoue, Y.H.、Taguchi, O.、Nishida, Y.、Matsukage, A. 和 Yamaguchi, M.：“人类骨髓增生异常果蝇同源物的表征/

Hirose, F. Yamaguchi, M. and Matsukage, A.: "Targeted expression of the DNA binding domain of DRE-binding factor, a Drosophila transcription factor, attenuates DNA replication of the salivary gland and eye imaginal disc."Mol. Cell. Biol.. 19. 6y020-6028 (

Hirose, F. Yamaguchi, M. 和 Matsukage, A.：“DRE 结合因子（一种果蝇转录因子）的 DNA 结合域的靶向表达可减弱唾液腺和眼成象盘的 DNA 复制。”

Izumi, M.et al.: "Transcription of the catalytic 180-kDa subunit gene of・・・"Biochem.Biophys.Acta.. 1492. 341-352 (2000)

Izumi, M.等人：“……的催化 180-kDa 亚基基因的转录”Biochem.Biophys.Acta.. 1492. 341-352 (2000)