Murawska Magdalena, Brehm Alexander
Institut für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität Marburg, Marburg, Germany.
Methods Mol Biol. 2012;809:267-77. doi: 10.1007/978-1-61779-376-9_18.
Gene transcription is a complex process that involves a large number of proteins. These proteins can be brought to their target genes by a variety of different mechanisms: many transcription factors interact with specific DNA sequences in promoters or enhancers, several epigenetic regulators bind histones bearing specific modifications, elongation factors and some RNA processing factors bind to the transcribing RNA polymerase, and other factors interact directly with nascent transcripts or noncoding RNA. Immunostaining of Drosophila polytene chromosomes allows the genome-wide localization of factors involved at different stages of transcriptional regulation. In this chapter, we present protocols that adapt the general technique to probe different recruitment mechanisms employed by these factors, including specific interactions with phosphorylated RNA polymerase II and RNA-mediated chromatin associations.
基因转录是一个复杂的过程，涉及大量蛋白质。这些蛋白质可通过多种不同机制被带到其靶基因：许多转录因子与启动子或增强子中的特定DNA序列相互作用，一些表观遗传调节因子与带有特定修饰的组蛋白结合，延伸因子和一些RNA加工因子与正在转录的RNA聚合酶结合，其他因子则直接与新生转录本或非编码RNA相互作用。果蝇多线染色体的免疫染色可实现全基因组范围内参与转录调控不同阶段的因子定位。在本章中，我们介绍了一些方案，这些方案对一般技术进行了调整，以探究这些因子所采用的不同募集机制，包括与磷酸化RNA聚合酶II的特异性相互作用以及RNA介导的染色质关联。
