Dynamics of RNA polymerase II and elongation factor Spt4/5 recruitment during activator-dependent transcription.

ABSTRACT: In eukaryotes, RNA polymerase II (RNApII) transcribes messenger RNA from template DNA. Decades of experiments have identified the proteins needed for transcription activation, initiation complex assembly, and productive elongation. However, the dynamics of recruitment of these proteins to transcription complexes, and of the transitions between these steps, are poorly understood. We used multiwavelength single-molecule fluorescence microscopy to directly image and quantitate these dynamics in a budding yeast nuclear extract that reconstitutes activator-dependent transcription in vitro. A strong activator (Gal4-VP16) greatly stimulated reversible binding of individual RNApII molecules to template DNA. Binding of labeled elongation factor Spt4/5 to DNA typically followed RNApII binding, was N

SUBMITTER: Rosen GA 

PROVIDER: S-EPMC7768755 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature