Unknown

Dataset Information

0

The displacement of the σ70 finger in initial transcription is highly heterogeneous and promoter-dependent.


ABSTRACT: Most bacterial sigma factors (σ) contain a highly conserved structural module, the 'σ-finger', which forms a loop that protrudes towards the RNA polymerase active centre in the open complex and has been implicated in pre-organization of template DNA, abortive initiation of short RNAs, initiation pausing, and promoter escape. Here, we introduce a novel single-molecule FRET (smFRET) assay to monitor σ-finger motions during transcription initiation and promoter escape. By performing real-time smFRET measurements, we determine that for all promoters studied, displacement occurs before promoter escape and can occur either before or after a clash with the extending RNA. We show that the kinetics of σ-finger displacement are highly dependent on the promoter, with implications for transcription kinetics and regulation. Analogous mechanisms may operate in the similar modules present across all kingdoms of life.

SUBMITTER: Wang A 

PROVIDER: S-EPMC12445709 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature

altmetric image

Publications

The displacement of the σ70 finger in initial transcription is highly heterogeneous and promoter-dependent.

Wang Anna A   Fletcher Andrew A   Mukherjee Pratip P   Grainger David C DC   Mazumder Abhishek A   Kapanidis Achillefs N AN  

Nucleic acids research 20250901 17


Most bacterial sigma factors (σ) contain a highly conserved structural module, the 'σ-finger', which forms a loop that protrudes towards the RNA polymerase active centre in the open complex and has been implicated in pre-organization of template DNA, abortive initiation of short RNAs, initiation pausing, and promoter escape. Here, we introduce a novel single-molecule FRET (smFRET) assay to monitor σ-finger motions during transcription initiation and promoter escape. By performing real-time smFRE  ...[more]

Similar Datasets

| S-EPMC3632114 | biostudies-literature
| S-EPMC5001585 | biostudies-literature
| S-EPMC4725480 | biostudies-literature
| S-EPMC7510298 | biostudies-literature
| S-EPMC10786286 | biostudies-literature