Pyocin S5 Import into Pseudomonas aeruginosa Reveals a Generic Mode of Bacteriocin Transport.

MBio
Hannah Michaela BehrensColin Kleanthous

Abstract

Pyocin S5 (PyoS5) is a potent protein bacteriocin that eradicates the human pathogen Pseudomonas aeruginosa in animal infection models, but its import mechanism is poorly understood. Here, using crystallography, biophysical and biochemical analyses, and live-cell imaging, we define the entry process of PyoS5 and reveal links to the transport mechanisms of other bacteriocins. In addition to its C-terminal pore-forming domain, elongated PyoS5 comprises two novel tandemly repeated kinked 3-helix bundle domains that structure-based alignments identify as key import domains in other pyocins. The central domain binds the lipid-bound common polysaccharide antigen, allowing the pyocin to accumulate on the cell surface. The N-terminal domain binds the ferric pyochelin transporter FptA while its associated disordered region binds the inner membrane protein TonB1, which together drive import of the bacteriocin across the outer membrane. Finally, we identify the minimal requirements for sensitizing Escherichia coli toward PyoS5, as well as other pyocins, and suggest that a generic pathway likely underpins the import of all TonB-dependent bacteriocins across the outer membrane of Gram-negative bacteria.IMPORTANCE Bacteriocins are toxic poly...Continue Reading

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Citations

Oct 13, 2020·Frontiers in Microbiology·William M RooneyDaniel Walker
Nov 7, 2020·Current Opinion in Structural Biology·Anna C RatliffHerve Celia
Jul 3, 2020·Journal of the American Chemical Society·Sejeong LeeHagan Bayley
Jun 19, 2021·The Journal of Antimicrobial Chemotherapy·Anne SixDaniel Walker
Aug 1, 2021·Nature Communications·Nicholas G HousdenColin Kleanthous
Sep 22, 2021·MBio·Ruth Cohen-KhaitColin Kleanthous

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Methods Mentioned

BETA
X-ray
light scattering
circular dichroism
isothermal titration calorimetry
chip
surface plasmon resonance
size
size exclusion chromatography
light-scattering

Software Mentioned

Staraniso
Buster
Phaser
Dials
MicroCal LLC Origin
CRYSOL
CCP4MG
BLASTn
AutoProc
ExPASy ProtParam

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