Spectrometric Nucleic Acid-Associated Protein ID

描述（由申请人提供） 微阵列技术的出现首次允许评估在特定压力、药物或毒物反应中发生的全基因组转录变化的能力。响应特定刺激而发生的基因变化波通过转录因子在特定启动子处的组装来传递。因此，将环境刺激与基因表达变化相结合的密码的关键组成部分是激活和失活转录因子在被诱导或抑制的启动子处的合成、修饰和聚集。对遗传网络中信息流的这一关键组成部分的研究受到了限制，因为难以识别和表征与表达受到调节的基因的完整调控区相互作用的蛋白质。虽然在鉴定哪些转录因子能够激活特定基因构建体方面已经取得了很大进展，但是关于哪些因子激活完整染色体中的实际基因以及当与完整基因相互作用时这些因子的次级修饰的直接知识是有限的。 为了促进对这些问题的研究，研究人员建议开发一种新技术，该技术将允许质谱鉴定与特定基因启动子相关的蛋白质。 如果成功，这项新技术有可能大大提高研究人员识别和表征蛋白质组装的能力，这些蛋白质组装在外部刺激后与特定启动子相互作用，从而为解开遗传网络的结构和组织做出重大贡献。研究人员建议用原核模型开发和验证这种方法，然后将该技术应用于调节形成环境敏感性生殖系统核心部分的具有良好特征的真核启动子。 具体而言，该技术将被应用到识别和表征的蛋白质相互作用的小鼠LhB启动子时，诱导脉冲激素管理。该基因具有良好表征的三级基因启动子，其需要诱导和合成其它转录因子以用于其诱导。研究人员将利用这种新开发的技术来鉴定在脉冲式GnRH诱导过程中与完整的LhB启动子结合的转录因子。

DESCRIPTION (provided by applicant) The advent of microarray technology has allowed, for the first time, the ability to assess the genome-wide changes in transcription that occur in response to specific stress, drugs or toxicants. The waves of gene changes that occur in response to a specific stimulus are transmitted by the assembly of transcription factors at specific promoters. Thus a key component of the code which couples environmental stimuli to changes in gene expression is the synthesis, modification and aggregation of activating and inactivating transcription factors at the promoters which are induced or suppressed. Studies of this crucial component of information flow in genetic networks have been limited by the difficulty to identify and characterize the proteins that interact with intact regulatory regions of genes whose expression is modulated. While great advances have been made in identifying which transcription factors are capable of activating particular gene constructs, direct knowledge is limited about which factors activate the actual gene in an intact chromosome and the secondary modification of those factors when interacting with the intact gene. In order to facilitate studies into these questions, the investigators propose to develop a new technique that will allow mass spectrometric identification of the proteins associated with specific gene promoters. If successful, this new technique has the potential to vastly improve the researchers' capacity to identify and characterize the protein assembly that interacts with specific promoters following an external stimulus and thereby significantly contribute to unraveling the structure and organization of genetic networks. The investigators propose to develop and validate this approach with a prokaryotic model, and then to apply the technique to the modulation of a well characterized eukaryotic promoter that forms a central part of the environmentally sensitive reproductive system. Specifically, the technique will be applied to the identification and characterization of proteins that interact with the mouse LhB promoter when induced by pulsatile hormone administration. This gene has a well characterized tertiary gene promoter that requires induction and synthesis of other transcription factors for its induction. The researchers will utilize this newly developed technique to identify the transcription factors that bind the intact LhB promoter during pulsatile GnRH induction.