Roles for HCF-1 in cell proliferation/differentiation

描述(申请人提供)：为了深入了解HCF-1在细胞周期控制中的作用，将结合生化和遗传学方法研究HCF-1与Myc/Max/Mad转录因子网络之间的相互作用，Myc/Max/Mad转录因子网络在控制细胞增殖、分化和凋亡方面发挥核心作用。其具体目的是：(1)探讨HCF-1-Mad的相互作用；(2)确定HCF-1-Mad的相互作用是否影响Mad的转录特性；(3)研究HCF-1-Mad的相互作用对细胞增殖和分化的影响。具体地说，免疫沉淀将被用来分析HCF-1与Mad的关联，以及这种关联是否受到其他蛋白质的影响，如Max、Myc和Sin3。电泳迁移率改变分析将用于检测HCF-1-Mad复合体的DNA结合活性，体内转录活性分析将提供关于HCF-1在调节Mad活性中的作用的信息。小干扰RNA将被用来耗尽内源性Mad，并研究在HCF-1结合中有缺陷的Mad突变形式的活性。这些研究将揭示HCF-1和Myc-Max-Mad网络在细胞增殖、分化和癌症进展中的生物学作用。

DESCRIPTION (provided by applicant): To gain insight into the roles of HCF-1 in cell cycle control, biochemical and genetic approaches will be combined to study the interaction between HCF-1 and the Myc/Max/Mad transcription factor network that plays a central role in controlling cellular proliferation, differentiation, and apoptosis. The specific aims are: (1) To probe HCF-1-Mad interactions; (2) To determine whether HCF-1-Mad interactions affect the transcription properties of Mad; and (3) To study the influence of HCF-1-Mad interaction on cell proliferation and differentiation. Specifically, immunoprecipitation will be used to analyze the association of HCF-1 with Mad, and whether this association is affected by other proteins, such as Max, Myc, and Sin3. Electrophoretic mobility shift assays will be used to examine the DNA-binding activity of the HCF-1-Mad complex, and in vivo transcription activity assays will provide information about roles of HCF-1 in modulating the activity of Mad. Small interfering RNAs will be used to deplete endogenous Mad and study the activities of mutant forms of Mad that are defective in HCF-1 association. These studies will reveal the biological roles of HCF-1 and Myc-Max-Mad network in cell proliferation, differentiation, and cancer progression.