Genome wide extracellular signal-regulated kinase (ERK) pathway components interaction with human genome

Several recent studies, including ours, show that most components of signal transduction cascades including the extracellular signal-regulated protein kinase (ERK), that once were thought to act predominantly in cytoplasm, are in fact recruited to chromatin and are integral components of transcriptional complexes. However, their distribution along whole genome remains uncovered. Here we investigate genome wide recruitment of the ERK pathway components using chromatin immunoprecipitations (ChIP) followed by deep sequencing, ChIP-Seq. Hela-S3 cells were starved for 48 hours (control) and stimulated with EGF at concentration of 100ng/mL for 20 minutes. Cells were fixed with formaldehyde, chromatin isolated and subjected to ChIP reaction. Two technical replicates of ChIP reaction followed by sequencing were performed.

包括本团队在内的多项近期研究表明，曾被认为主要在细胞质中发挥功能的多数信号转导级联反应组分，包括细胞外信号调节激酶（extracellular signal-regulated protein kinase, ERK），实际上会被招募至染色质，并作为转录复合物的核心组成部分。然而，此类组分在全基因组范围内的分布特征仍未被揭示。本研究采用染色质免疫共沉淀（chromatin immunoprecipitation, ChIP）结合深度测序技术（即ChIP-Seq），对ERK通路组分的全基因组招募情况展开探究。实验以HeLa-S3细胞为研究模型，将其血清饥饿培养48小时作为对照组，随后以100ng/mL浓度的表皮生长因子（epidermal growth factor, EGF）刺激20分钟。之后使用甲醛固定细胞，分离染色质并开展ChIP反应，共完成两组技术重复的ChIP实验及后续测序。