Crystal structure of <i>Aquifex aeolicus</i> σ^N bound to promoter DNA and the structure of σ^N-holoenzyme.

Abstract

The bacterial σ factors confer promoter specificity to the RNA polymerase (RNAP). One alternative σ factor, σ^N, is unique in its structure and functional mechanism, forming transcriptionally inactive promoter complexes that require activation by specialized AAA⁺ ATPases. We report a 3.4-Å resolution X-ray crystal structure of a σ^N fragment in complex with its cognate promoter DNA, revealing the molecular details of promoter recognition by σ^N The structure allowed us to build and refine an improved σ^N-holoenzyme model based on previously published 3.8-Å resolution X-ray data. The improved σ^N-holoenzyme model reveals a conserved interdomain interface within σ^N that, when disrupted by mutations, leads to transcription activity without activator intervention (so-called bypass mutants). Thus, the structure and stability of this interdomain interface are crucial for the role of σ^N in blocking transcription activity and in maintaining the activator sensitivity of σ^N.