# Single-stranded DNA drives σ subunit loading onto mycobacterial RNA polymerase to unlock initiation-competent conformations

## TL;DR

This study reveals how single-stranded DNA helps the σ subunit load onto mycobacterial RNA polymerase, enabling transcription initiation.

## Contribution

The study identifies a novel mechanism where ssDNA guides RNA polymerase maturation and σ subunit regulation.

## Key findings

- −10 ssDNA induces σ subunit loading onto RNA polymerase channels.
- ssDNA triggers RNAP clamp closure and unswiveling necessary for transcription initiation.
- σ subunit acts as a regulator of RNA polymerase conformational dynamics.

## Abstract

Initiation of transcription requires the formation of the “open” promoter complex (RPo). For this, the σ subunit of bacterial RNA polymerase (RNAP) binds to the nontemplate strand of the −10 element sequence of promoters and nucleates DNA unwinding. This is accompanied by a cascade of conformational changes on RNAP, the exact mechanics of which remains elusive. Here, using single-molecule Förster resonance energy transfer and cryo-electron microscopy, we explored the conformational landscape of RNAP from the human pathogen Mycobacterium tuberculosis upon binding to a single-stranded DNA (ssDNA) fragment that includes the −10 element sequence (−10 ssDNA). We found that like the transcription activator RNAP-binding protein A, −10 ssDNA induced σ subunit loading onto the DNA/RNA channels of RNAP. This triggered RNAP clamp closure and unswiveling that are required for RPo formation and RNA synthesis initiation. Our results reveal a mechanism of ssDNA-guided RNAP maturation and identify the σ subunit as a regulator of RNAP conformational dynamics.

Graphical Abstract