Genome-wide chromatin binding of transcriptional corepressor Topless-related 1 in Arabidopsis. Genome-wide chromatin binding of transcriptional corepressor Topless-related 1 in Arabidopsis

Plant Topless-related 1 (TPR1), belonging to a family of transcriptional corepressors found across eukaryotes, contributes to immunity signaling in Arabidopsis thaliana and wild tobacco. We used chromatin immunoprecipitation and sequencing (ChIP-seq) of Arabidopsis TPR1-GFP expressing transgenic lines to characterize genome-wide TPR1-chromatin associations. Overall design: We performed ChIP-seq experiments with α-GFP antibodies in three independent biological replicates using leaves of six-week-old pTPR1:TPR1-GFP in Col-0, a line with constitutive immunity ,and pTPR1:TPR1-GFP in eds1-2 plants lacking constitutive immunity. The pTPR1:TPR1-HA #3 Col-0 line served as a negative control as it is also autoimmune and accumulates similar TPR1 transcript levels as the TPR1-GFP line. Leaf material of 6-week-old plants was used in all three independent biological replicates.

植物Topless相关蛋白1（Plant Topless-related 1, TPR1）属于一类在真核生物中广泛分布的转录共抑制因子家族，参与拟南芥（Arabidopsis thaliana）与野生烟草的免疫信号传导过程。本研究通过对表达TPR1-GFP的拟南芥转基因株系开展染色质免疫共沉淀测序（chromatin immunoprecipitation and sequencing, ChIP-seq），以解析全基因组范围内TPR1与染色质的结合关联。实验整体设计：我们采用抗GFP抗体（α-GFP），以哥伦比亚生态型Col-0背景下携带pTPR1:TPR1-GFP的组成型免疫株系、以及eds1-2突变体背景下携带pTPR1:TPR1-GFP的无组成型免疫株系的六周龄叶片为材料，完成了三次独立生物学重复的ChIP-seq实验。其中，pTPR1:TPR1-HA #3 Col-0株系被设为阴性对照：该株系同样具备自身免疫表型，且TPR1转录本的积累水平与TPR1-GFP株系基本一致。所有三次独立生物学重复均使用六周龄植株的叶片材料。