TAP TAG PULL DOWN OF NOVEL PROTEINS INVOLVED IN TGM SILENCING

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在转录水平上有许多类型的调控元件。某些类型的调控元件导致转录的正增强和其他负调控或沉默。在酿酒酵母中，一些被研究和了解最多的类型是端粒附近的沉默和沉默的交配位点。我们的实验室在酵母中发现了一种新的沉默形式，它涉及到RNA聚合酶III转录tRNA基因。导致其他类型沉默缓解的基因缺失已被证明对TGM沉默没有影响，然而我们实验室之前的研究已经发现了缓解这种现象的新蛋白质。然而，这些蛋白质都没有被证明具有任何沉默能力或相互作用。我们认为，这些蛋白质中的一种必须与直接导致tRNA基因座沉默的蛋白质相互作用。我们已经获得了这些蛋白质的双重亲和版本，并将进行下拉分析，然后使用质谱学来识别新的蛋白质相互作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There are many types of regulatory elements at the transcriptional level. Some types of regulatory elements cause positive enhancement of transcription and other negative regulation or ?silencing.? In Saccaromyces cerevisiae some of the most studied and understood types are silencing near telomeres and silent mating loci. Our lab has identified a novel form of silencing in yeast which involves the transcription of tRNA genes by RNA polymerase III. The transcription of pol III tRNA genes has been shown to cause the inhibition of certain RNA polymerase II genes. Deletion of genes shown to cause alleviation of other types of silencing has shown to have no affect on TGM silencing, however previous studies in our lab has uncovered novel proteins that alleviates this phenomena. However, none of these proteins has been shown to possess any silencing ability or to interact with each other. It is our belief that one of these proteins must be interacting with proteins that directly lead to silencing that the tRNA loci. We have acquired duel affinity versions of these proteins and will do pull downs and then use Mass spectrometry to identify novel protein interactions.