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PMID:11160803

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Citation

Hennequin, C, Porcheray, F, Waligora-Dupriet, A, Collignon, A, Barc, M, Bourlioux, P and Karjalainen, T (2001) GroEL (Hsp60) of Clostridium difficile is involved in cell adherence. Microbiology (Reading, Engl.) 147:87-96

Abstract

Previous results have demonstrated that adherence of Clostridium difficile to tissue culture cells is augmented by various stresses; this study focussed on whether the GroEL heat shock protein is implicated in this process. The 1940 bp groESL operon of C. difficile was isolated by PCR. The 1623 bp groEL gene is highly conserved between various C. difficile isolates as determined by RFLP-PCR and DNA sequencing, and the operon is present in one copy on the bacterial chromosome. The 58 kDa GroEL protein was expressed in Escherichia coli in fusion with glutathione S:-transferase and the fusion protein was purified from IPTG-induced bacterial lysates by affinity chromatography on glutathione-Sepharose. A polyclonal, monospecific antiserum was obtained for GroEL which established by immunoelectron microscopy, indirect immunofluorescence and immunoblot analysis that GroEL is released extracellularly after heat shock and can be surface associated. Cell fractionation experiments suggest that GroEL is predominantly cytoplasmic and membrane bound. GroEL-specific antibodies as well as the purified protein partially inhibited C. difficile cell attachment and expression of the protein was induced by cell contact, suggesting a role for GroEL in cell adherence.

Links

PubMed

Keywords

Amino Acid Sequence; Animals; Bacterial Adhesion/physiology; Base Sequence; Cercopithecus aethiops; Chaperonin 60/genetics; Chaperonin 60/isolation & purification; Chaperonin 60/metabolism; Clostridium difficile/pathogenicity; Clostridium difficile/physiology; Enterocolitis, Pseudomembranous/microbiology; Gene Dosage; Humans; Microscopy, Immunoelectron; Molecular Sequence Data; Operon; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Sequence Analysis, DNA; Vero Cells

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

CLODI:Q9FA27

located_in

GO:0009274: peptidoglycan-based cell wall

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:CH60

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:CH60

located_in

GO:0009274: peptidoglycan-based cell wall

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:CH60

located_in

GO:0005615: extracellular space

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:CH60

located_in

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:CH60

located_in

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CLODI:Q9FA27

part_of

GO:0005618: cell wall

ECO:0000314: direct assay evidence used in manual assertion

C

exists_during:(GO:0009408)

Seeded From UniProt

complete

PEPDI:CH60

GO:0044650: adhesion of symbiont to host cell

ECO:0000314:

P

Figure 5 shows the adhesion of C. difficile to Vero cells in the absence and presence of anti-GroEL; adhesion decreases when anti-GroEL is present indicating that GroEL is involved in the attachment of C. difficile to cells.

complete
CACAO 7979

PEPDI:CH60

involved_in

GO:0044650: adhesion of symbiont to host cell

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

PEPDI:CH60

part_of

GO:0005615: extracellular space

ECO:0000314: direct assay evidence used in manual assertion

C

exists_during:(GO:0009408)

Seeded From UniProt

complete

PEPDI:CH60

part_of

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PEPDI:CH60

part_of

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PEPDI:CH60

part_of

GO:0005618: cell wall

ECO:0000314: direct assay evidence used in manual assertion

C

exists_during:(GO:0009408)

Seeded From UniProt

complete

PEPDI:CH60

part_of

GO:0009986: cell surface

ECO:0000314: direct assay evidence used in manual assertion

C

exists_during:(GO:0009408)

Seeded From UniProt

complete


See also

References

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