TY - JOUR

T1 - An enzyme moonlights as a chaperone to control virulence of Pseudomonas aeruginosa

AU - Borgert, Sebastian Roman

AU - Henke, Steffi

AU - Witzgall, Florian

AU - Schmelz, Stefan

AU - zur Lage, Susanne

AU - Hotop, Sven Kevin

AU - Stephen, Steffi

AU - Lübken, Dennis

AU - Krüger, Jonas

AU - Gomez, Nicolas Oswaldo

AU - van Ham, Marco

AU - Jänsch, Lothar

AU - Kalesse, Markus

AU - Pich, Andreas

AU - Brönstrup, Mark

AU - Häussler, Susanne

AU - Blankenfeldt, Wulf

N1 - Funding Information: We are grateful to the staff of beamline P11 at the PETRAIII synchrotron (DESY campus Hamburg, Germany) for letting us use their facilities. Peer Lukat is acknowledged for help with data collection. S.R.B. received a scholarship by the Life Sciences Foundation and was supported by the HZI International Graduate School for Infection Research (GS-FIRE). S.He. is a member of the PROCOMPAS graduate school supported by the Deutsche Forschungsgemeinschaft (281361126/GRK2223).

PY - 2022/12/1

Y1 - 2022/12/1

N2 - Pseudomonas aeruginosa is a major cause of nosocomial infections and also leads to severe exacerbations in cystic fibrosis or chronic obstructive pulmonary disease. Three intertwined quorum sensing systems control virulence of P. aeruginosa, with the rhl circuit playing the leading role in late and chronic infections. The majority of traits controlled by rhl transcription factor RhlR depend on PqsE, a dispensable thioesterase in Pseudomonas Quinolone Signal (PQS) biosynthesis that interferes with RhlR through an enigmatic mechanism likely involving direct interaction of both proteins. Here we show that PqsE and RhlR form a 2:2 protein complex that, together with RhlR agonist N-butanoyl-L-homoserine lactone (C4-HSL), solubilizes RhlR and thereby renders the otherwise insoluble transcription factor active. We determine crystal structures of the complex and identify residues essential for the interaction. To corroborate the chaperone-like activity of PqsE, we design stability-optimized variants of RhlR that bypass the need for C4-HSL and PqsE in activating PqsE/RhlR-controlled processes of P. aeruginosa. Together, our data provide insight into the unique regulatory role of PqsE and lay groundwork for developing new P. aeruginosa-specific pharmaceuticals.

AB - Pseudomonas aeruginosa is a major cause of nosocomial infections and also leads to severe exacerbations in cystic fibrosis or chronic obstructive pulmonary disease. Three intertwined quorum sensing systems control virulence of P. aeruginosa, with the rhl circuit playing the leading role in late and chronic infections. The majority of traits controlled by rhl transcription factor RhlR depend on PqsE, a dispensable thioesterase in Pseudomonas Quinolone Signal (PQS) biosynthesis that interferes with RhlR through an enigmatic mechanism likely involving direct interaction of both proteins. Here we show that PqsE and RhlR form a 2:2 protein complex that, together with RhlR agonist N-butanoyl-L-homoserine lactone (C4-HSL), solubilizes RhlR and thereby renders the otherwise insoluble transcription factor active. We determine crystal structures of the complex and identify residues essential for the interaction. To corroborate the chaperone-like activity of PqsE, we design stability-optimized variants of RhlR that bypass the need for C4-HSL and PqsE in activating PqsE/RhlR-controlled processes of P. aeruginosa. Together, our data provide insight into the unique regulatory role of PqsE and lay groundwork for developing new P. aeruginosa-specific pharmaceuticals.

UR - http://www.scopus.com/inward/record.url?scp=85143184742&partnerID=8YFLogxK

U2 - 10.21203/rs.3.rs-1489597/v1

DO - 10.21203/rs.3.rs-1489597/v1

M3 - Article

C2 - 36456567

AN - SCOPUS:85143184742

VL - 13

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 7402

ER -