ADENOVIRUS E1A PROTEIN AND THE SWI/SNF COMPLEX

描述：(改编自调查人员摘要)：腺病毒E1a 蛋白质诱导细胞在培养过程中的分化和转化 因此是研究细胞转化的一个很好的模型。E1a 结合特定的细胞蛋白，包括视网膜母细胞瘤蛋白(Rb)和 部分通过改变染色质来控制转录的P300/CBP蛋白 结构。E1a抑制染色质重塑复合体的功能 Swi/Snf，它与Rb蛋白相互作用，可能与p300相互作用。具体目标1 建议定义和表征哺乳动物的SWI/SNF复合体 它在转录调控中的作用。这将决定如何 复合体在转录水平抑制c-fos的表达。结构性的 伴随着c-fos基因活性抑制而来的变化将是 特征，并将开发检测方法来测量物理关联 在体内用特定的哺乳动物启动子研究SWI/SNF。研究将表现出 E1a对SWI/SNF过程的影响。《特定目标2》将使用酵母菌 用遗传学方法了解E1a对SWI/SNF复合体的抑制作用。酵母菌 将确定与E1a相互作用的蛋白质及其对E1a的影响 活动将被确定。随后将进行类似的研究。 哺乳动物细胞使用从酵母系统中获得的信息。

DESCRIPTION:(Adapted from the Investigator's abstract): The adenovirus E1A protein induces imortalization and transformation of cells in culture and is therefore an excellent model for the study of cellular transformation. E1A binds specific cell proteins including the retinoblastoma protein (Rb) and p300/CBP proteins that control transcription, in part, by altering chromatin structure. E1A inhibits the function of the chromatin remodeling complex SWI/SNF, which interacts with Rb protein and possibly p300. Specific aim 1 proposes to define and characterize the mammalian SWI/SNF complex with respect to its role in regulation of transcription. It will be determined how the complex inhibits c-fos expression at the transcriptional level. The structural changes that accompany the inhibition of c-fos gene activity will be characterized, and assays will be developed to measure the physical association of SWI/SNF with specific mammalian promoters in vivo. Studies will characterize the effects of E1A on SWI/SNF processes. Specific aim 2 will use a yeast genetic approach to understand E1A inhibition of the SWI/SNF complex. The yeast proteins that interact with E1A will be identified and their influence on E1A activity will be determined. Similar studies will then be conducted in mammalian cells using the information gained from the yeast system.