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Structure paper

TitleStructural Basis for Virulence Activation of Francisella tularensis.
Journal, issue, pagesMol Cell, Vol. 81, Issue 1, Page 139-152.e10, Year 2021
Publish dateJan 7, 2021
AuthorsBrady A Travis / Kathryn M Ramsey / Samantha M Prezioso / Thomas Tallo / Jamie M Wandzilak / Allen Hsu / Mario Borgnia / Alberto Bartesaghi / Simon L Dove / Richard G Brennan / Maria A Schumacher /
PubMed AbstractThe bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, ...The bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, and the stress signal, ppGpp. MglA-SspA assembles with the σ-associated RNAP holoenzyme (RNAPσ), forming a virulence-specialized polymerase. These factors activate Francisella pathogenicity island (FPI) gene expression, which is required for virulence, but the mechanism is unknown. Here we report FtRNAPσ-promoter-DNA, FtRNAPσ-(MglA-SspA)-promoter DNA, and FtRNAPσ-(MglA-SspA)-ppGpp-PigR-promoter DNA cryo-EM structures. Structural and genetic analyses show MglA-SspA facilitates σ binding to DNA to regulate virulence and virulence-enhancing genes. Our Escherichia coli RNAPσhomodimeric EcSspA structure suggests this is a general SspA-transcription regulation mechanism. Strikingly, our FtRNAPσ-(MglA-SspA)-ppGpp-PigR-DNA structure reveals ppGpp binding to MglA-SspA tethers PigR to promoters. PigR in turn recruits FtRNAP αCTDs to DNA UP elements. Thus, these studies unveil a unique mechanism for Ft pathogenesis involving a virulence-specialized RNAP that employs two (MglA-SspA)-based strategies to activate virulence genes.
External linksMol Cell / PubMed:33217319 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.95 - 4.43 Å
Structure data

EMDB-21850, PDB-6wmp:
F. tularensis RNAPs70-iglA DNA complex
Method: EM (single particle) / Resolution: 2.98 Å

EMDB-21851, PDB-6wmr:
F. tularensis RNAPs70-(MglA-SspA)-iglA DNA complex
Method: EM (single particle) / Resolution: 3.46 Å

EMDB-21852, PDB-6wmt:
F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA complex
Method: EM (single particle) / Resolution: 4.43 Å

EMDB-21853, PDB-6wmu:
E. coli RNAPs70-SspA-gadA DNA complex
Method: EM (single particle) / Resolution: 3.18 Å

PDB-6weg:
Structure of Ft (MglA-SspA)-ppGpp-PigR peptide complex
Method: X-RAY DIFFRACTION / Resolution: 2.95 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-G4P:
GUANOSINE-5',3'-TETRAPHOSPHATE / Guanosine pentaphosphate

ChemComp-ZN:
Unknown entry

Source
  • francisella tularensis subsp. holarctica lvs (bacteria)
  • francisella tularensis subsp. holarctica (strain lvs) (bacteria)
  • escherichia coli (E. coli)
  • escherichia coli ta054 (bacteria)
  • francisella tularensis subsp. tularensis (strain schu s4 / schu 4) (bacteria)
  • francisella tularensis (bacteria)
KeywordsTRANSCRIPTION / Francisella tularensis / bioweapon / MglA-SspA / PigR / ppGpp / RNA polymerase complex

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