Abstract:
MYC is a transcription factor playing multiple functions both in physiological and pathological settings. Biochemical characterizations, combined with the analyses of MYC chromatin binding, have shown that its pleiotropic activity depends on the chromatin context and its protein-protein interactions with different cofactors. In order to determine the contribution of MYC in a certain biological condition, it would be relevant to analyze the concomitant binding of MYC and its associated proteins, in relationship to the chromatin environment. To this end, we here provide a simple method to parallel map the genome-wide binding of MYC-associated proteins, together with the chromatin profiling of multiple histone modifications. We detail the procedure to perform high-throughput ChIP-seq (HT-ChIP-seq) with a variety of biological samples. In addition, we describe simple bioinformatic steps to determine the distribution of MYC binding with respect to the chromatin context and the association of its cofactors. The described approach will permit the reproducible characterization of MYC activity in different biological contexts.
摘要:
MYC是在生理和病理环境中发挥多种功能的转录因子。生化特征,结合MYC染色质结合的分析,已表明其多效活性取决于染色质环境及其与不同辅因子的蛋白质-蛋白质相互作用。为了确定MYC在一定生物学条件下的贡献,这将与分析MYC及其相关蛋白的伴随结合有关,与染色质环境有关。为此,我们在这里提供了一种简单的方法来平行绘制MYC相关蛋白的全基因组结合图,以及多种组蛋白修饰的染色质谱。我们详细介绍了对各种生物样品执行高通量ChIP-seq(HT-ChIP-seq)的过程。此外,我们描述了简单的生物信息学步骤来确定MYC结合相对于染色质环境的分布及其辅因子的关联。所描述的方法将允许在不同生物学背景下对MYC活性进行可再现的表征。