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TitleBidirectional pilus processing in the Tad pilus system motor CpaF.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 6635, Year 2024
Publish dateAug 5, 2024
AuthorsMichael Hohl / Emma J Banks / Max P Manley / Tung B K Le / Harry H Low /
PubMed AbstractThe bacterial tight adherence pilus system (TadPS) assembles surface pili essential for adhesion and colonisation in many human pathogens. Pilus dynamics are powered by the ATPase CpaF (TadA), which ...The bacterial tight adherence pilus system (TadPS) assembles surface pili essential for adhesion and colonisation in many human pathogens. Pilus dynamics are powered by the ATPase CpaF (TadA), which drives extension and retraction cycles in Caulobacter crescentus through an unknown mechanism. Here we use cryogenic electron microscopy and cell-based light microscopy to characterise CpaF mechanism. We show that CpaF assembles into a hexamer with C2 symmetry in different nucleotide states. Nucleotide cycling occurs through an intra-subunit clamp-like mechanism that promotes sequential conformational changes between subunits. Moreover, a comparison of the active sites with different nucleotides bound suggests a mechanism for bidirectional motion. Conserved CpaF residues, predicted to interact with platform proteins CpaG (TadB) and CpaH (TadC), are mutated in vivo to establish their role in pilus processing. Our findings provide a model for how CpaF drives TadPS pilus dynamics and have broad implications for how other ancient type 4 filament family members power pilus assembly.
External linksNat Commun / PubMed:39103374 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 4.0 Å
Structure data

19209
EMDB entry, No image

EMDB-19209, PDB-8rjf:
TadA/CpaF with ADP
Method: EM (single particle) / Resolution: 3.1 Å

19275
EMDB entry, No image

EMDB-19275, PDB-8rkd:
TadA/CpaF with AMPPNP
Method: EM (single particle) / Resolution: 3.8 Å

19279
EMDB entry, No image

EMDB-19279, PDB-8rkl:
TadA/CpaF nucleotide free
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-EPE:
4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID / pH buffer*YM

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

Source
  • caulobacter vibrioides na1000 (bacteria)
KeywordsMOTOR PROTEIN / pilus / secretion / ATPase / Apo

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