Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK

Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK

ABSTRACT The DnaK (Hsp70) protein is an essential ATP‐dependent chaperone foldase and holdase found in most organisms. In this study, combining multiple experimental approaches we determined FliC as major interaction partner of DnaK in the opportunistic bacterial pathogen Pseudomonas aeruginosa. Imp...

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Main Authors: Gabriella Molinari, Sara S. Ribeiro, Katrin Müller, Benjamin E. Mayer, Manfred Rohde, Alejandro Arce‐Rodriguez, Juan José Vargas‐Guerrero, Albert Avetisyan, Josef Wissing, Werner Tegge, Lothar Jänsch, Mark Brönstrup, Antoine Danchin, Martina Jahn, Kenneth N. Timmis, Simon Ebbinghaus, Dieter Jahn, José Manuel Borrero‐de Acuña
Format: Article
Language:English
Published: Wiley 2025-02-01
Series:Microbial Biotechnology
Subjects:
DnaK
flagellum
FliC
Hsp70
Pseudomonas aeruginosa
Online Access:https://doi.org/10.1111/1751-7915.70096
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Summary:ABSTRACT The DnaK (Hsp70) protein is an essential ATP‐dependent chaperone foldase and holdase found in most organisms. In this study, combining multiple experimental approaches we determined FliC as major interaction partner of DnaK in the opportunistic bacterial pathogen Pseudomonas aeruginosa. Implementing immunofluorescence microscopy and electron microscopy techniques DnaK was found extracellularly associated to the assembled filament in a regular pattern. dnaK repression led to intracellular FliC accumulation and motility impairment, highlighting DnaK essentiality for FliC export and flagellum assembly. SPOT–membrane peptide arrays coupled with artificial intelligence analyses suggested a highly dynamic DnaK–FliC interaction landscape involving multiple domains and transient complexes formation. Remarkably, in vitro fast relaxation imaging (FReI) experiments mimicking ATP‐deprived extracellular environment conditions exhibited DnaK ATP‐independent holdase activity, regardless of its co‐chaperone DnaJ and its nucleotide exchange factor GrpE. We present a model for the DnaK‐FliC interactions involving dynamic states throughout the flagellum assembly stages. These results expand the classical view of DnaK chaperone functioning and introduce a new participant in the Pseudomonas flagellar system, an important trait for bacterial colonisation and virulence.
ISSN:1751-7915

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