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TitleAntiparallel stacking of Csu pili drives Acinetobacter baumannii 3D biofilm assembly.
Journal, issue, pagesNat Commun, Year 2026
Publish dateFeb 7, 2026
AuthorsHenri Malmi / Natalia Pakharukova / Bindusmita Paul / Minna Tuittila / Irfan Ahmad / Stefan David Knight / Bernt Eric Uhlin / Debnath Ghosal / Anton V Zavialov /
PubMed AbstractMany Gram-negative nosocomial pathogens rely on adhesive filaments, known as archaic chaperone-usher pili, to establish stress- and drug-resistant, multi-layered biofilms. Here, we uncover the ...Many Gram-negative nosocomial pathogens rely on adhesive filaments, known as archaic chaperone-usher pili, to establish stress- and drug-resistant, multi-layered biofilms. Here, we uncover the mechanism by which these pili build three-dimensional (3D) biofilm architectures. In situ analyses of Acinetobacter baumannii biofilms using electron microscopy (EM) reveal an extensive network of ultrathin, flat stacks of archaic Csu pili interconnecting bacterial cells in 3D space. Cryo-EM structures of a single native pilus, pilus pairs, and two types of multi-pilus stacks show that the pili pack into antiparallel sheets, with their rods connected laterally by junctions at their zigzag corners. This antiparallel arrangement ensures that contacts form primarily between pili from interacting cells rather than pili from the same cell. With a remarkably short helical repeat, archaic chaperone-usher pili spontaneously establish a high density of junctions that determines the biofilm's 3D architecture. Our findings may help develop new therapies against multidrug-resistant bacterial infections by targeting pilus-pilus interactions.
External linksNat Commun / PubMed:41654547
MethodsEM (helical sym.) / EM (single particle)
Resolution3.28 - 8.91 Å
Structure data

52587

9i37

EMDB-52587, PDB-9i37:
Csu pilus rod purified from Acinetobacter baumannii
Method: EM (helical sym.) / Resolution: 3.28 Å

52600

9i3m

EMDB-52600, PDB-9i3m:
Csu pilus rod antiparallel pair
Method: EM (single particle) / Resolution: 8.91 Å

52601

9i3n

EMDB-52601, PDB-9i3n:
Csu pilus rod type 1 stack
Method: EM (single particle) / Resolution: 7.58 Å

52602

9i3o

EMDB-52602, PDB-9i3o:
Csu pilus rod type 2 stack
Method: EM (single particle) / Resolution: 4.7 Å

Source
  • acinetobacter baumannii (bacteria)
KeywordsCELL ADHESION / pili / fimbriae / Acinetobacter baumannii pili / chaperone-usher pathway / archaic chaperone-usher pili / biofilm / 3D biofilm / adhesion / pathogenesis / pilus antiparallel binding junction

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