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TitleCyclic di-AMP traps proton-coupled K transporters of the KUP family in an inward-occluded conformation.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 3683, Year 2023
Publish dateJun 21, 2023
AuthorsMichael F Fuss / Jan-Philip Wieferig / Robin A Corey / Yvonne Hellmich / Igor Tascón / Joana S Sousa / Phillip J Stansfeld / Janet Vonck / Inga Hänelt /
PubMed AbstractCyclic di-AMP is the only known essential second messenger in bacteria and archaea, regulating different proteins indispensable for numerous physiological processes. In particular, it controls ...Cyclic di-AMP is the only known essential second messenger in bacteria and archaea, regulating different proteins indispensable for numerous physiological processes. In particular, it controls various potassium and osmolyte transporters involved in osmoregulation. In Bacillus subtilis, the K/H symporter KimA of the KUP family is inactivated by c-di-AMP. KimA sustains survival at potassium limitation at low external pH by mediating potassium ion uptake. However, at elevated intracellular K concentrations, further K accumulation would be toxic. In this study, we reveal the molecular basis of how c-di-AMP binding inhibits KimA. We report cryo-EM structures of KimA with bound c-di-AMP in detergent solution and reconstituted in amphipols. By combining structural data with functional assays and molecular dynamics simulations we reveal how c-di-AMP modulates transport. We show that an intracellular loop in the transmembrane domain interacts with c-di-AMP bound to the adjacent cytosolic domain. This reduces the mobility of transmembrane helices at the cytosolic side of the K binding site and therefore traps KimA in an inward-occluded conformation.
External linksNat Commun / PubMed:37344476 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 3.8 Å
Structure data

15894

8b70

EMDB-15894, PDB-8b70:
KimA from B. subtilis with nucleotide second-messenger c-di-AMP bound
Method: EM (single particle) / Resolution: 3.3 Å

15895

8b71

EMDB-15895, PDB-8b71:
Upright KimA dimer with bound c-di-AMP from B. subtilis
Method: EM (single particle) / Resolution: 3.8 Å

Chemicals

ChemComp-2BA:
(2R,3R,3aS,5R,7aR,9R,10R,10aS,12R,14aR)-2,9-bis(6-amino-9H-purin-9-yl)octahydro-2H,7H-difuro[3,2-d:3',2'-j][1,3,7,9,2,8

ChemComp-LMT:
DODECYL-BETA-D-MALTOSIDE / detergent*YM

ChemComp-K:
Unknown entry

Source
  • bacillus subtilis (bacteria)
KeywordsMEMBRANE PROTEIN / potassium importer / second-messenger / c-di-AMP / second messenger

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