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TitleStructural basis of transcriptional activation by the Mycobacterium tuberculosis intrinsic antibiotic-resistance transcription factor WhiB7.
Journal, issue, pagesMol Cell, Vol. 81, Issue 14, Page 2875-22886.e5, Year 2021
Publish dateJul 15, 2021
AuthorsMirjana Lilic / Seth A Darst / Elizabeth A Campbell /
PubMed AbstractIn pathogenic mycobacteria, transcriptional responses to antibiotics result in induced antibiotic resistance. WhiB7 belongs to the Actinobacteria-specific family of Fe-S-containing transcription ...In pathogenic mycobacteria, transcriptional responses to antibiotics result in induced antibiotic resistance. WhiB7 belongs to the Actinobacteria-specific family of Fe-S-containing transcription factors and plays a crucial role in inducible antibiotic resistance in mycobacteria. Here, we present cryoelectron microscopy structures of Mycobacterium tuberculosis transcriptional regulatory complexes comprising RNA polymerase σ-holoenzyme, global regulators CarD and RbpA, and WhiB7, bound to a WhiB7-regulated promoter. The structures reveal how WhiB7 interacts with σ-holoenzyme while simultaneously interacting with an AT-rich sequence element via its AT-hook. Evidently, AT-hooks, rare elements in bacteria yet prevalent in eukaryotes, bind to target AT-rich DNA sequences similarly to the nuclear chromosome binding proteins. Unexpectedly, a subset of particles contained a WhiB7-stabilized closed promoter complex, revealing this intermediate's structure, and we apply kinetic modeling and biochemical assays to rationalize how WhiB7 activates transcription. Altogether, our work presents a comprehensive view of how WhiB7 serves to activate gene expression leading to antibiotic resistance.
External linksMol Cell / PubMed:34171296 / PubMed Central
MethodsEM (single particle)
Resolution2.74 - 3.38 Å
Structure data

22886

7kif

EMDB-22886, PDB-7kif:
Mycobacterium tuberculosis WT RNAP transcription open promoter complex with WhiB7 transcription factor
Method: EM (single particle) / Resolution: 2.94 Å

22887

7kim

EMDB-22887, PDB-7kim:
Mycobacterium tuberculosis WT RNAP transcription closed promoter complex with WhiB7 transcription factor
Method: EM (single particle) / Resolution: 3.38 Å

22888

7kin

EMDB-22888, PDB-7kin:
Mycobacterium tuberculosis WT RNAP transcription open promoter complex with WhiB7 promoter
Method: EM (single particle) / Resolution: 2.74 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-SF4:
IRON/SULFUR CLUSTER / Iron–sulfur cluster

Source
  • mycobacterium tuberculosis (bacteria)
KeywordsTRANSCRIPTION / TRANSFERASE/DNA / RNA polymerase / Transcription factor / Mycobacterium tuberculosis / Iron cluster / TRANSFERASE-DNA complex

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