Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [I]

The formation of heterochromatin at HML, HMR, and telomeres in Saccharomyces cerevisiae involves two main steps: Recruitment of Sir proteins to silencers and their spread throughout the silenced domain. For the following datasets, we created a fusion protein between the heterochromatin protein Sir3 and the non-site-specific bacterial adenine methyltransferase M.EcoGII, with or without a 3xV5 epitope at the C-terminus. We performed ChIP-seq experiments (immunoprecipitated Sir3-M.EcoGII-3xV5) and MeDIP-seq experiments (immunoprecipitated m6A methylated DNA). We also used a temperature-sensitive allele of SIR3 (sir3-8) fused to M.ECOGII to induce m6A methylation for MeDIP-seq. 32 samples total (24 in initial submission, 8 more in resubmission). Input and IP data for each experiment are included.

酿酒酵母（Saccharomyces cerevisiae）中HML、HMR及端粒处异染色质的形成主要包含两个核心步骤：将Sir蛋白招募至沉默子位点，并使其在沉默结构域内扩散。针对本数据集，我们构建了异染色质蛋白Sir3与非位点特异性细菌腺嘌呤甲基转移酶M.EcoGII的融合蛋白，该融合蛋白的C端可携带或不携带3xV5表位。我们开展了染色质免疫共沉淀测序（ChIP-seq）实验（免疫沉淀靶标为Sir3-M.EcoGII-3xV5融合蛋白）与甲基化DNA免疫沉淀测序（MeDIP-seq）实验（免疫沉淀N6-甲基腺嘌呤（m6A）修饰的DNA）。此外，我们还使用了与M.ECOGII融合的SIR3温度敏感等位基因sir3-8，以诱导用于MeDIP-seq实验的m6A甲基化修饰。本数据集共计32个样本（初始提交版本含24个，补充提交新增8个），所有实验均配套提供输入样本与免疫沉淀样本的数据。