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TitleThe molecular basis of protein toxin HicA-dependent binding of the protein antitoxin HicB to DNA.
Journal, issue, pagesJ Biol Chem, Vol. 293, Issue 50, Page 19429-19440, Year 2018
Publish dateDec 14, 2018
AuthorsAshley J Winter / Christopher Williams / Michail N Isupov / Hannah Crocker / Mariya Gromova / Philip Marsh / Oliver J Wilkinson / Mark S Dillingham / Nicholas J Harmer / Richard W Titball / Matthew P Crump /
PubMed AbstractToxin-antitoxin (TA) systems are present in many bacteria and play important roles in bacterial growth, physiology, and pathogenicity. Those that are best studied are the type II TA systems, in which ...Toxin-antitoxin (TA) systems are present in many bacteria and play important roles in bacterial growth, physiology, and pathogenicity. Those that are best studied are the type II TA systems, in which both toxins and antitoxins are proteins. The HicAB system is one of the prototypic TA systems, found in many bacterial species. Complex interactions between the protein toxin (HicA), the protein antitoxin (HicB), and the DNA upstream of the encoding genes regulate the activity of this system, but few structural details are available about how HicA destabilizes the HicB-DNA complex. Here, we determined the X-ray structures of HicB and the HicAB complex to 1.8 and 2.5 Å resolution, respectively, and characterized their DNA interactions. This revealed that HicB forms a tetramer and HicA and HicB form a heterooctameric complex that involves structural reorganization of the C-terminal (DNA-binding) region of HicB. Our observations indicated that HicA has a profound impact on binding of HicB to DNA sequences upstream of in a stoichiometric-dependent way. At low ratios of HicA:HicB, there was no effect on DNA binding, but at higher ratios, the affinity for DNA declined cooperatively, driving dissociation of the HicA:HicB:DNA complex. These results reveal the structural mechanisms by which HicA de-represses the HicB-DNA complex.
External linksJ Biol Chem / PubMed:30337369 / PubMed Central
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution1.56 - 2.49 Å
Structure data

SASDD45:
Type II toxin-antitoxin system HicB family antitoxin - HicB protein
Method: SAXS/SANS

SASDD55:
Type II toxin-antitoxin system HicB family antitoxin - HicB protein - bound to addiction module toxin, HicA
Method: SAXS/SANS

PDB-6g1c:
Crystal structure of the N-terminal domain of Burkholderia Pseudomallei antitoxin HicB
Method: X-RAY DIFFRACTION / Resolution: 1.56 Å

PDB-6g1n:
Crystal structure of the Burkholderia Pseudomallei antitoxin HicB
Method: X-RAY DIFFRACTION / Resolution: 1.85 Å

PDB-6g26:
The crystal structure of the Burkholderia pseudomallei HicAB complex
Method: X-RAY DIFFRACTION / Resolution: 2.49 Å

Chemicals

ChemComp-HOH:
WATER

ChemComp-CL:
Unknown entry

ChemComp-GOL:
GLYCEROL

ChemComp-SO4:
SULFATE ION

ChemComp-EDO:
1,2-ETHANEDIOL

ChemComp-PGE:
TRIETHYLENE GLYCOL

Source
  • burkholderia pseudomallei (bacteria)
  • burkholderia pseudomallei k96243 (bacteria)
KeywordsANTITOXIN / N-terminal domain of the antitoxin HicB which acts as an inhibitor to HicA / The antitoxin HicB which acts as an inhibitor to HicA

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