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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| Format: | Article |
| Language: | English |
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Wiley
2025-02-01
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| Series: | Microbial Biotechnology |
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| Online Access: | https://doi.org/10.1111/1751-7915.70096 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Gabriella Molinari |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-636ee9b3de8e4431added2eefb7ea5ac |
| institution | OA Journals |
| issn | 1751-7915 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj-art-636ee9b3de8e4431added2eefb7ea5ac2025-08-20T02:01:58ZengWileyMicrobial Biotechnology1751-79152025-02-01182n/an/a10.1111/1751-7915.70096Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaKGabriella Molinari0Sara S. Ribeiro1Katrin Müller2Benjamin E. Mayer3Manfred Rohde4Alejandro Arce‐Rodriguez5Juan José Vargas‐Guerrero6Albert Avetisyan7Josef Wissing8Werner Tegge9Lothar Jänsch10Mark Brönstrup11Antoine Danchin12Martina Jahn13Kenneth N. Timmis14Simon Ebbinghaus15Dieter Jahn16José Manuel Borrero‐de Acuña17Central Facility for Microscopy Helmholtz Centre for Infection Research (HZI) Braunschweig GermanyInstitute of Physical and Theoretical Chemistry Technische Universität Braunschweig Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyComputational Biology and Simulation Technische Universität Darmstadt Darmstadt GermanyCentral Facility for Microscopy Helmholtz Centre for Infection Research (HZI) Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyInstitute of Physical and Theoretical Chemistry Technische Universität Braunschweig Braunschweig GermanyDepartment Cellular Proteome Research Helmholtz Centre for Infection Research (HZI) Braunschweig GermanyDepartment of Chemical Biology Helmholtz Centre for Infection Research (HZI) Braunschweig GermanyDepartment Cellular Proteome Research Helmholtz Centre for Infection Research (HZI) Braunschweig GermanyDepartment of Chemical Biology Helmholtz Centre for Infection Research (HZI) Braunschweig GermanySchool of Biomedical Sciences, Li KaShing Faculty of Medicine The University of Hong Kong SAR Hong Kong ChinaInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyInstitute of Physical and Theoretical Chemistry Technische Universität Braunschweig Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyInstitute of Microbiology Technische Universität Braunschweig Braunschweig GermanyABSTRACT 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.https://doi.org/10.1111/1751-7915.70096DnaKflagellumFliCHsp70Pseudomonas aeruginosa |
| spellingShingle | 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 Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK Microbial Biotechnology DnaK flagellum FliC Hsp70 Pseudomonas aeruginosa |
| title | Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK |
| title_full | Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK |
| title_fullStr | Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK |
| title_full_unstemmed | Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK |
| title_short | Multiple Chaperone DnaK–FliC Flagellin Interactions are Required for Pseudomonas aeruginosa Flagellum Assembly and Indicate a New Function for DnaK |
| title_sort | multiple chaperone dnak flic flagellin interactions are required for pseudomonas aeruginosa flagellum assembly and indicate a new function for dnak |
| topic | DnaK flagellum FliC Hsp70 Pseudomonas aeruginosa |
| url | https://doi.org/10.1111/1751-7915.70096 |
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