THE EFFECTS OF SUB2 PROTEIN ON GENE SILENCING IN S CEREVISIA

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。 通过实时荧光定量聚合酶链式反应和染色质免疫沉淀法对端粒沉默的检测，确定端粒沉默是否可以与S在转录延伸中的作用分开。我们将评估是否能够将Sub2定位在当Sub2过度表达时上调的基因的启动子上。我们还将通过生化分析来确定Sub2是否可以与特定的转录因子结合。三个转录因子已经在细菌中通过共纯化实验被证实为Sub2p相互作用因子。然而，一个更严格的标准是在酵母而不是细菌中重建这种相互作用。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.