Abstract
This chapter focuses on methods to study the early steps in transcript synthesis such as initiation, escape commitment, and promoter escape. These approaches are used to determine how different factors affect specific steps in early RNA synthesis. Each of the steps in early transcription has the potential to be influenced by transcriptional activators and/or general transcription factors such as TFIIE and TFIIH. For example, initiation and postinitiation events, promoter proximal pausing, promoter clearance, and transcript elongation have been shown to be targets of transcriptional regulators. In addition, TFIIH contains two ATP-dependent helicases thought to function during early steps in transcription. The TFIIE and TFIIH increase the fraction of complexes that produce full-length RNA by acting at a specific step in early transcription—namely, escape commitment.
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