TSS identification of Rap1-regulated transcripts by 5' end RNA sequencing

Eukaryotic cells utilize several mechanisms to ensure that expression of aberrant non-coding RNAs is limited. Gene looping, chromatin modification or remodeling, and RNA surveillance contribute to ensure the fidelity of transcription and limit non-coding transcripts. We have identified that the transcription factor Rap1 is critical for limiting the expression of aberrant RNAs, particularly near the highly expressed ribosomal protein genes. Here we utilize a 5’ end RNA sequencing protocol to identify transcript TSSs at single nucleotide resolution in wild-type and Rap1 auxin-inducible degron strains of Saccharomyces cerevisiae.真核细胞拥有多种调控机制，以限制异常非编码RNA（non-coding RNA）的表达。基因环化、染色质修饰与重塑以及RNA监视（RNA surveillance）通路，可保障转录的保真度，并限制非编码转录本的生成。我们已证实，转录因子Rap1在限制异常RNA的表达中发挥关键作用，尤其在高表达的核糖体蛋白基因邻近区域。本研究采用5’端RNA测序（5’ end RNA sequencing）实验方案，在酿酒酵母（Saccharomyces cerevisiae）的野生型菌株与Rap1生长素诱导降解菌株中，以单核苷酸分辨率实现转录起始位点（Transcription Start Sites, TSSs）的精准鉴定。