TRANSCRIPTIONAL COACTIVATORS IN HEMATOPOIESIS

描述：(改编自调查人员摘要)抄本 Ets-1因子在造血细胞功能中发挥重要作用 目前还不清楚它能激活转录。一些转录因子 使用转录共激活物CREB结合蛋白(CBP)和 介导TNA聚合酶II(POL II)反式激活的相关p300。多么 CBP/p300介导反式激活尚不清楚，但它结合基础 转录因子，并与组蛋白乙酰转移酶(HAT)和 蛋白激酶活性可能在激活POL II中起作用。 CBP相关的HAT活性在造血中的重要性 急性髓系T(8；16)(p11；p13)易位 一种白血病，假定HAT与CBP融合。这样做的长期目标是 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及其方式 这些相互作用的扰动会导致疾病状态。这 提案的重点是假设ETS-1调节POL II 通过结合CBP/p300的特定区域进行转录。ETS-1活动可以 受调节Ets-1：CBP/p300相互作用的影响，或受调节 与CBP/p300相关的因子和酶活性。具体目标一 将测试ETS-1是否需要与特定的CBP/P300域交互 Ets-1转录活性。申请者将定义CBP/p300 Ets-1与体内外结合的充要序列 并将其与体内的反式激活功能相关联。特定目标 II：将使用缺失和点突变来从功能上定义 在体内和体外与CBP/p300相互作用所需的Ets-1序列。它 将确定这些ETS-1序列是否必要，以及 足以在体内发挥反式激活功能。特定目标III将 确定ETS-1活性是否与CBP/p300相关因子和 酶活性。研究人员将测量HAT，蛋白激酶 和POL II活性以及普遍存在的转录因子 Ets-1：CBP/p300复合体及其与Ets-1反式激活的关系 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.