Dissociation between the critical role of ClpB of Francisella tularensis for the heat shock response and the DnaK interaction and its important role for efficient type VI secretion and bacterial virulence.

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  • Additional Information
    • Source:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101238921 Publication Model: eCollection Cited Medium: Internet ISSN: 1553-7374 (Electronic) Linking ISSN: 15537366 NLM ISO Abbreviation: PLoS Pathog Subsets: MEDLINE
    • Publication Information:
      Original Publication: San Francisco, CA : Public Library of Science, c2005-
    • Subject Terms:
      Endopeptidase Clp/*metabolism ; Francisella tularensis/*physiology ; HSP70 Heat-Shock Proteins/*metabolism; Animals ; Bacterial Proteins/metabolism ; Endopeptidase Clp/genetics ; Escherichia coli/metabolism ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Female ; Francisella tularensis/genetics ; Francisella tularensis/metabolism ; Francisella tularensis/pathogenicity ; HSP70 Heat-Shock Proteins/genetics ; Heat-Shock Proteins/genetics ; Heat-Shock Proteins/metabolism ; Heat-Shock Response ; Mice ; Mice, Inbred C57BL ; Molecular Chaperones/metabolism ; Molecular Dynamics Simulation ; Type VI Secretion Systems/metabolism ; Virulence/physiology
    • Abstract:
      Francisella tularensis, a highly infectious, intracellular bacterium possesses an atypical type VI secretion system (T6SS), which is essential for its virulence. The chaperone ClpB, a member of the Hsp100/Clp family, is involved in Francisella T6SS disassembly and type VI secretion (T6S) is impaired in its absence. We asked if the role of ClpB for T6S was related to its prototypical role for the disaggregation activity. The latter is dependent on its interaction with the DnaK/Hsp70 chaperone system. Key residues of the ClpB-DnaK interaction were identified by molecular dynamic simulation and verified by targeted mutagenesis. Using such targeted mutants, it was found that the F. novicida ClpB-DnaK interaction was dispensable for T6S, intracellular replication, and virulence in a mouse model, although essential for handling of heat shock. Moreover, by mutagenesis of key amino acids of the Walker A, Walker B, and Arginine finger motifs of each of the two Nucleotide-Binding Domains, their critical roles for heat shock, T6S, intracellular replication, and virulence were identified. In contrast, the N-terminus was dispensable for heat shock, but required for T6S, intracellular replication, and virulence. Complementation of the ΔclpB mutant with a chimeric F. novicida ClpB expressing the N-terminal of Escherichia coli, led to reconstitution of the wild-type phenotype. Collectively, the data demonstrate that the ClpB-DnaK interaction does not contribute to T6S, whereas the N-terminal and NBD domains displayed critical roles for T6S and virulence.
      Competing Interests: The authors have declared that no competing interests exist.
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    • Accession Number:
      0 (Bacterial Proteins)
      0 (Escherichia coli Proteins)
      0 (HSP70 Heat-Shock Proteins)
      0 (Heat-Shock Proteins)
      0 (Molecular Chaperones)
      0 (Type VI Secretion Systems)
      EC 3.4.21.92 (Endopeptidase Clp)
      EC 3.6.1.- (dnaK protein, E coli)
      EC 3.6.1.3 (ClpB protein, E coli)
    • Publication Date:
      Date Created: 20200411 Date Completed: 20200804 Latest Revision: 20200804
    • Publication Date:
      20231215
    • Accession Number:
      PMC7182274
    • Accession Number:
      10.1371/journal.ppat.1008466
    • Accession Number:
      32275693