DNA origami-based single-molecule force spectroscopy elucidates RNA Polymerase III pre-initiation complex stability

• DOI: 10.1038/s41467-020-16702-x
• Publication Date: 2020-06-05T10:04:22Z
• Authors: Kevin Kramm; Tim Schröder; Jerome Gouge; Andrés Manuel Vera; Kapil Gupta; Florian B. Heiss; Tim Liedl; Christoph Engel; Imre Berger; Alessandro Vannini; Philip Tinnefeld; Dina Grohmann
• Keywords: Force Spectroscopy; Transcription; DNA clamp; RNA polymerase II

Abstract The TATA-binding protein (TBP) and a transcription factor (TF) IIB-like are important constituents of all eukaryotic initiation complexes. reason for the emergence strict requirement additional Bdp1 in RNA polymerase (RNAP) III system, however, remained elusive. A poorly studied aspect this context is effect DNA strain arising from compaction transcriptional activity on complex formation. We made use origami-based force clamp to follow assembly human complexes RNAP II systems at single-molecule level under piconewton forces. demonstrate that TBP-DNA force-sensitive TFIIB sufficient stabilise TBP strained promoter. In contrast, pivotal component ensures stable anchoring factors, thus itself, system. Thereby, we offer an explanation crucial role high output III.