Growth control of hemaotpoietic stem cells by the collaboration of transcription factors, cell-cycle regulators, and tumor suppressors

通过转录因子、细胞周期调节剂和肿瘤抑制因子的协作控制造血干细胞的生长

• 批准号: 18591037
• 负责人: MINEGISHI Naoko
• 金额: $ 2.43万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591037/
• 关键词: Transcriptional Regulation; Transplantation and regenerative medicine; Stem cell; Hematopoiesis; Transcription factor; 移植・再生医療; 造血幹細胞; 細胞周期; 細胞増殖; アポトーシス; リン酸化; 蛋白質分解; 静止期

Cell-cycle regulation of hematopoietic stem cells is of interest because understanding of this mechanism appears to facilitate the development of ex-vivo methods to manipulate various stem cells. GATA2 is an indispensable transcription factor regulating proliferation and quiescence of hematopoietic stem cells. Here we show that GATA-2 and cyclin dependent kinase (Cdk)/Cyclin complexes bidirectionally control their expressions. In leukemic and normal immature hematopoietic cells, GATA-2 expression is high in G0/G1 and S/G2 phase, and low in early S and G2/M phase. Immunoprecipitation/immunoblotting analysis demonstrated phosphorylation of GATA-2 at Cdk-consensus motifs, S/T_0P_<+1>, and interaction of GATA2 with Cdk2/Cyclin A2. Cdk2/Cyclin A2, Cdk2/Cyclin El, or Cdk4/Cyclin D1 directly phosphorylated immunoprecipitated GATA-2 in vitro. Treatment with proteasome inhibitor MG132, or Cdk inhibitor Roscovitine modulated cell cycle-dependent oscillation of GATA-2 expression. Mutations in Cdk … More -consensus motifs of GATA-2 exhibited altered expression profiles of green fluorescent protein-GATA2 domain fusion proteins. Therefore, Cdk/Cyclin phosphorylated GATA-2 and regulated its proteasome-dependent degradation. On the other hand, while GATA-2, Cyclin El and Cyclin D2 were expressed in embryonic day 10.5 wild type placenta, CyclinEl and CyclinD2 mRNA expressions were significantly lower in placentas with hypomorphic Gata2 loci. ChIP experiments of synchronized leukemic cells revealed GATA-2 binding to CyclinEl and CyclinD2 gene at late G1 phase, and detachment of GATA-2 in early S phase. Transient GATA-2 binding in late G1 phase was accompanied by RNA polymerase II binding to these genes and concomitant CyclinEl and CyclinD2 mRNA expressions, and was suppressed by 1-hour treatment with Roscovitine. Consequently, GATA2 bound and regulated transcription of CyclinD2 and CyclinEl gene, while Cyclin D2, Cyclin El and other Cyclin, forming complexes with Cdk, phohsphorylated GATA-2 and modulate stability of this factor. This bidirectional regulation between GATA-2 and Cdk/Cyclins might determine the cell-cycle characteristics of GATA-2 expressing hematopoietic cells. Less

造血干细胞的细胞周期调节是感兴趣的，因为对这种机制的理解似乎有助于开发前体的方法来操纵各种干细胞。 GATA2是必不可少的转录因子调节和造血干细胞的静止。在这里，我们表明GATA-2和细胞周期蛋白依赖性激酶（CDK）/细胞周期蛋白复合物双向控制其表达。在白血病和正常未成熟的造血细胞中，G0/G1和S/G2相的GATA-2表达很高，在早期S和G2/M期间较低。免疫沉淀/免疫印迹分析表明，在CDK-CONSENSUS基序中GATA-2，S/T_0P _ <+1>以及GATA2与CDK2/Cyclin A2的相互作用。在体外，CDK2/Cyclin A2，CDK2/Cyclin EL或CDK4/Cyclin D1直接磷酸化的免疫沉淀GATA-2。用蛋白酶体抑制剂MG132或CDK抑制剂Roscovitine调制细胞周期依赖性GATA-2表达的振荡处理。 CDK中的突变…GATA-2的更多传感器基序暴露于绿色荧光蛋白-GATA2结构域融合蛋白的表达谱。因此，CDK/细胞周期蛋白磷酸化的GATA-2并调节其蛋白酶体依赖性降解。另一方面，在胚胎第10.5天野生型胎盘中表达了GATA-2，细胞周期蛋白EL和细胞周期蛋白D2，而胎盘中的Cyclinel和Cyclind2 mRNA表达明显较低，胎盘含量明显较低。同步的白血病细胞的芯片实验显示GATA-2在G1后期与Cyclinel和Cyclind2基因结合，并在早期S阶段脱离GATA-2。 G1后期的瞬时GATA-2结合伴随着RNA聚合酶II与这些基因的结合，并伴有cyclinel和Cyclind2 mRNA表达，并通过1小时的Roscovitine抑制。因此，GATA2结合了Cyclind2和Cyclinel基因的转录和调节的转录，而细胞周期蛋白D2，Cyclin EL和其他细胞周期蛋白则与CDK形成复合物，PhohSphyphorySpated GATA-2并调节该因素的稳定性。 GATA-2和CDK/细胞周期蛋白之间的这种双向调节可能会决定GATA-2表达造血细胞的细胞周期特征。较少的

项目成果

転写因子GATA-2とCyclin/Cdk系との双方向性制御

转录因子GATA-2和Cyclin/Cdk系统的双向调控

• 发表时间: 2007
• 作者: Satoh K;et. al.;Tada H.;Satoh K;Tada H;Harigae H;峯岸 直子
• 通讯作者: 峯岸 直子

Regulation of adipocyte differentiation of bone marrow stromal cells by transcription factor GATA-2.

• DOI: 10.1016/j.bbrc.2007.10.031
• 发表时间: 2007-12
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Y. Okitsu;Shinichirou Takahashi;N. Minegishi;J. Kameoka;M. Kaku;Masayuki Yamamoto;Takeshi Sasaki;H. Harigae

Phenotypic changes of hematopoietic stem cells and stromal cells by aberrant expression of GATA-2; possible role for the development of aplastic Anemia

GATA-2 异常表达导致造血干细胞和基质细胞表型变化；

• 发表时间: 2006
• 作者: Mukai HY;et. al.;Okitsu Y ed. al.
• 通讯作者: Okitsu Y ed. al.

Phenotypic changes of hematopoietic stem cells and stromal cells by aberrant expression of GATA-2; possible role for the development of aplastic Anemia.

GATA-2 异常表达导致造血干细胞和基质细胞表型变化；

• 发表时间: 2006
• 作者: Mukai HY ;et. al.;Okitsu Y
• 通讯作者: Okitsu Y

個々の細胞レベルの遺伝子発現解析によって解明された転写制御と細胞周期制御の造血前駆細胞におけるクロストーク

通过个体细胞水平的基因表达分析阐明造血祖细胞转录调控和细胞周期调控之间的串扰

• 发表时间: 2007
• 作者: Satoh K;et. al.;Tada H.;Satoh K;Tada H;Harigae H;峯岸 直子;Minegishi N ed. al.;峯岸 直子;峯岸 直子;峯岸 直子
• 通讯作者: 峯岸 直子