THE EFFECTS OF SUB2 PROTEIN ON GENE SILENCING IN S CEREVISIA

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。使用实时PCR和染色质免疫沉淀分析来监测Sub 2的位置，以确定Sub 2 p对端粒沉默的影响是否可以与Sub 2在转录延伸中的作用分开。我们将评估是否可以将Sub 2定位在当Sub 2过表达时上调的基因的启动子处。我们还将确定是否Sub 2可以结合到特定的转录因子，使用生化测定。三个转录因子已经被验证为Sub 2 p相互作用使用共纯化测定在细菌中。然而，一个更严格的标准是在酵母中而不是在细菌中重现这种相互作用。我们目前正在进行这些实验。

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. Determine whether the effect of Sub2p on telomere silencing can be separated from Sub2's role in transcription elongation, using real-time PCR and chromatin immunoprecipitation assays to monitor the location of Sub2. We will assess whether we can locate Sub2 at the promoter of genes that are upregulated when Sub2 is overexpressed. We will also determine whether Sub2 can bind to specific transcription factors using biochemical assays. Three transcription factors have already been verified as Sub2p interactors using co-purification assays in bacteria. However, a more rigid standard would be to recreate this interaction in yeast rather than in bacteria. We are currently pursuing those experiments.