TRANSCRIPTIONAL COACTIVATORS IN HEMATOPOIESIS

描述：（改编自研究者摘要） Ets-1因子在造血细胞功能中起着重要作用，但如何发挥作用呢？ 它激活转录还不清楚。 许多转录因子 使用转录辅激活因子CREB结合蛋白（CBP）和 相关p300介导TNA聚合酶II（Pol II）的反式激活。 如何 CBP/p300介导的反式激活是未知的，但它结合基础 转录因子和相关的组蛋白乙酰转移酶（HAT）， 蛋白激酶活性，可能在激活Pol II中发挥作用。 的 CBP相关的HAT活性在造血中的重要性由 在急性髓系白血病中发现t（8;16）（p11;p13）易位 假定的HAT与CBP融合的白血病。 这一长期目标 建议是定义造血转录的机制， 因子通过与CBP/p300相互作用刺激基因表达，以及如何 这些相互作用的扰动导致疾病状态。 这 一项提案的重点是Ets-1调节Pol II的假设 通过结合CBP/p300的特定区域进行转录。 Ets-1活性可以 通过调节Ets-1：CBP/p300相互作用或通过调节 与CBP/p300相关的因子和酶活性。 具体目标I 将检测Ets-1是否需要与特定CBP/p300结构域相互作用 Ets-1转录活性。 申请人将定义CBP/p300 Ets-1体外和体内结合所必需和充分的序列 并将其与体内反式激活功能相关联。 具体目标 II：缺失和点诱变将用于功能性地定义突变。 Ets-1序列需要在体内和体外与CBP/p300相互作用。 它 将确定这些Ets-1序列是否必要， 足以在体内发挥反式激活功能。 具体目标III将 确定Ets-1活性是否与CBP/p300相关因子相关， 酶活性 研究人员将测量HAT，蛋白激酶 和Pol II活性以及存在于 Ets-1：CBP/p300复合物，并将其与Ets-1反式激活相关 潜力

DESCRIPTION: (Adapted from investigator's abstract) The transcription factor Ets-1 plays an important role in hematopoietic cell function, but how it activates transcription is unclear. A number of transcription factors use the transcription coactivators CREB binding protein (CBP) and the related p300 to mediate transactivation of TNA polymerase II (Pol II). How CBP/p300 mediates transactivation is unknown, but it binds basal transcription factors and has associated histone acetyltransferase (HAT) and protein kinase activities that may play a role in activating Pol II. The importance of CBP associated HAT activity in hematopoiesis is suggested by the t(8;16)(p11;p13) translocation found in a subset of acute myeloid leukemias where a putative HAT is fused to CBP. The long term goals of this proposal are to define the mechanisms by which hematopoietic transcription factors stimulate gene expression by interacting with CBP/p300 and how perturbation of these interactions results in a disease state. This proposal is focused on the hypothesis that Ets-1 regulates Pol II transcription by binding specific regions of CBP/p300. Ets-1 activity can be affected by modulating the Ets-1:CBP/p300 interaction, or by regulating factors and enzymatic activities associated with CBP/p300. Specific Aim I will test if interaction of Ets-1 with specific CBP/p300 domains is required for Ets-1 transcription activity. The applicant will define CBP/p300 sequences necessary and sufficient for Ets-1 binding in vitro and in vivo and will correlate this with transactivation function in vivo. Specific Aim II: Deletion and point mutagenesis will be used to functionally define the Ets-1 sequences required to interact in vivo and in vitro with CBP/p300. It will be determined whether these Ets-1 sequences are necessary and sufficient for transactivation function in vivo. Specific Aim III will determine if Ets-1 activity correlates with CBP/p300 associated factors and enzymatic activities. The investigators will measure HAT, protein kinase and Pol II activities and general transcription factors present in Ets-1:CBP/p300 complexes and correlate this with Ets-1 transactivation potential.