TAP TAG PULL DOWN OF NOVEL PROTEINS INVOLVED IN TGM SILENCING

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。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引起转录的正向增强和其他负向调控或沉默。在酿酒酵母菌中，一些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。我们的实验室已经在酵母中发现了一种新的沉默形式，它涉及RNA聚合酶III对tRNA基因的转录。pol III tRNA基因的转录已被证明可引起某些RNA聚合酶II基因的抑制。缺失导致其他类型沉默减轻的基因已被证明对TGM沉默没有影响，但我们实验室先前的研究发现了减轻这种现象的新蛋白质。然而，这些蛋白质都没有显示出具有任何沉默能力或相互作用。我们相信这些蛋白质中的一个一定与直接导致tRNA位点沉默的蛋白质相互作用。我们已经获得了这些蛋白质的双亲和版本，并将进行下拉，然后使用质谱法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.