Nucleosome positioning - Transient Depletion and Recovery of the Essential RSC Chromatin Remodeling Complex [MNase-Seq]

we use an induced degron system to study how rapid clearance of ATP-dependent chromatin remodelers affect nucleosome locations. 1. Screening eight remodelers 2. depletion of Sth1 leads to rapid and reversible fill-in of nucleosome-free regions at gene promoters. 3. changes at depletion and recovery do not depend on major chromatin reassembly during DNA replication. 4. RSC is crucial in maintaining open promoters of low expressed genes. RSC is not directly required for an acute transcriptional response. 5. Changes in nucleosome positions are tightly related to the specific TSS usage at promoters.本研究借助诱导型降解子系统（induced degron system），探究ATP依赖型染色质重塑因子（ATP-dependent chromatin remodelers）被快速清除后对核小体定位的影响。 1. 筛选八种染色质重塑因子 2. Sth1耗竭会导致基因启动子处的无核小体区域（nucleosome-free regions）被快速且可逆地填补 3. 耗竭与恢复阶段的染色质变化不依赖于DNA复制过程中的主要染色质重组装 4. RSC复合物对于维持低表达基因的开放启动子至关重要，且RSC并非急性转录反应（acute transcriptional response）的直接必需因子 5. 核小体位置的变化与启动子处特定转录起始位点（transcription start site, TSS）的使用情况紧密相关。