TY - JOUR
T1 - Moonlighting chaperone activity of the enzyme PqsE contributes to RhlR-controlled 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 - © 2022. The Author(s).
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.
KW - Virulence
KW - Pseudomonas aeruginosa/genetics
KW - Protein Folding
KW - Transcription Factors
UR - http://www.scopus.com/inward/record.url?scp=85143184742&partnerID=8YFLogxK
U2 - 10.1038/s41467-022-35030-w
DO - 10.1038/s41467-022-35030-w
M3 - Journal article
C2 - 36456567
SN - 2041-1722
VL - 13
SP - 7402
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 7402
ER -