Bidirectional pilus processing in the Tad pilus system motor CpaF

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Aug. 5, 2024

Abstract 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 retraction cycles Caulobacter crescentus through an unknown mechanism. Here we use cryogenic electron microscopy cell-based light to characterise We show that into a hexamer with C2 symmetry different nucleotide states. Nucleotide cycling occurs intra-subunit clamp-like mechanism promotes sequential conformational changes between subunits. Moreover, comparison of active sites nucleotides bound suggests bidirectional motion. Conserved residues, predicted interact platform proteins CpaG (TadB) CpaH (TadC), mutated vivo establish their role processing. Our findings provide model how TadPS have broad implications other ancient type 4 filament family members power assembly.

Language: Английский

---

PilY1 regulates the dynamic architecture of the type IV pilus machine in Pseudomonas aeruginosa

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Oct. 30, 2024

Language: Английский

---

Conformational changes in the motor ATPase CpaF facilitate a rotary mechanism of Tad pilus assembly

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: April 24, 2025

Abstract The type IV pilus family uses PilT/VirB11-like ATPases to rapidly assemble and disassemble pilin subunits. Among these, the tight adherence (Tad) performs both functions using a single bifunctional ATPase, CpaF. Here, we determine three conformationally distinct structures of CpaF hexamers with varying nucleotide occupancies by cryo-electron microscopy. Analysis these suggest ATP binding hydrolysis expand rotate hexamer pore clockwise while subsequent ADP release contracts ATPase. Truncation intrinsically disordered region in Caulobacter crescentus equally reduces extension retraction events observed fluorescence microscopy, but does not reduce ATPase activity. AlphaFold3 modeling suggests that other motors filament superfamily employ conserved secondary structural features engage their respective platform proteins. From data, propose clockwise, rotary mechanism catalysis right-handed, helical Tad pilus, process broadly applicable motor systems.

Language: Английский

---

Methyloraptor flagellatus gen. Nov., sp. nov., novel Ancalomicrobiaceae-affiliated facultatively methylotrophic bacteria that feed on methanotrophs of the genus Methylococcus

Systematic and Applied Microbiology, Journal Year: 2024, Volume and Issue: 48(1), P. 126565 - 126565

Published: Nov. 22, 2024

Language: Английский

---

Editorial overview: Rise of the bacterial nanomachines

Current Opinion in Microbiology, Journal Year: 2024, Volume and Issue: 80, P. 102515 - 102515

Published: July 21, 2024

Language: Английский

---

Citrus sinensis fruit peel phytochemicals inhibit Pseudomonas aeruginosa biofilms and suppress motility phenotypes by potentially targeting quorum sensing pathways

South African Journal of Botany, Journal Year: 2024, Volume and Issue: 174, P. 152 - 166

Published: Sept. 12, 2024

Language: Английский

---

Coordinated conformational changes in the Tad pilus ATPase CpaF facilitate a rotary mechanism of catalysis

Published: Oct. 28, 2024

ABSTRACT The type IV pilus family uses PilT-like ATPases to rapidly assemble and disassemble pilin subunits. Among them, the tight adherence (Tad) performs both functions using a single bifunctional ATPase, CpaF. How ATP catalysis by CpaF facilitates assembly disassembly of Tad remains unclear. Here, we determined electron cryo-microscopy structures in three distinct conformations nucleotide occupancies. Packing unit analyses revealed differences coordination active sites, as well synchronized domain movements during catalytic cycle. Alphafold3 modeling demonstrated that other motors within filament superfamily use similar binding interface engage their respective platform proteins. From these data, propose employs clockwise, rotary mechanism catalysis, which couples chemical energy mechanical force biogenesis pili, process broadly applicable motor systems.

Language: Английский

---

Antivirulence therapy: type IV pilus as a druggable target for bacterial infections

Medicinal Chemistry Research, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 4, 2024

Language: Английский

---

Building permits—control of type IV pilus assembly by PilB and its cofactors

Journal of Bacteriology, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 7, 2024

Many bacteria produce type IV pili (T4P), surfaced-exposed protein filaments that enable cells to interact with their environment and transition from planktonic surface-adapted states. T4P are dynamic, undergoing rapid cycles of filament extension retraction facilitated by a complex nanomachine powered cytoplasmic motor ATPases. Dedicated assembly motors drive the pilus fiber into extracellular space, but like any machine, this process is tightly organized. These coordinated various ligands binding partners, which control or optimize functional associations machinery before commit crucial first step building pilus. This review focuses on molecular mechanisms regulate function. We discuss secondary messenger-dependent transcriptional post-translational regulation acting both directly through effectors. also recent discoveries naturally occurring inhibitors as well alternative motor-dependent signaling pathways. Given important virulence factors for many bacterial pathogens, studying these regulatory systems will provide new insights T4P-dependent physiology efficient strategies disable them.

Language: Английский

---

Naturally-occurring, strain-specific defects in the retraction ofAcinetobacter baumanniitype IV pili promote biofilm formation

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 14, 2024

Abstract Type IV pili are helical filaments composed of protein subunits which produced by numerous taxa bacteria, including Acinetobacter . extended out from the cell extension enzyme complexes, extract membrane and insert them into base filament, but can also be retracted reverse rotation catalyzed a retraction enzyme. have diverse functions, some (twitching motility, DNA-uptake) require while others (host adhesion, bacterial aggregation) do not. International Clone I (IC-I) II (IC-II) strains, show variable phenotypes in assays type pilus-dependent functions. We this variation is result efficiency pilus between subtypes, that, differential balance retraction-dependent retraction-independent define subtypes based on sequence major subunit, PilA. In both naturally-occurring pilA variants IC-I IC-II groups isogenic strains complemented with or , subtype promotes greater twitching motility DNA-uptake biofilm formation showing reduced capacity for similar to retraction-deficient mutant consistent hypothesis that naturally deficient. Testing defect was sufficient increase level piliation surface, we compared yields T4P sheared surface found an background, complementation results levels PilA per than equivalent gene.

Language: Английский

---