High-resolution visualization by field emission scanning electron microscopy of Enterococcus faecalis surface proteins encoded by the pheromone-inducible conjugative plasmid pCF10.
Enterococcus faecalis can acquire antibiotic resistance and virulence genes by transfer of pheromone-inducible conjugative plasmids such as pCF10, which encodes tetracycline resistance. Two pCF10-encoded cell surface proteins, Sec10 and Asc10, have been previously shown to play an important role in the transfer of this plasmid. We used high-resolution, field emission scanning electron microscopy to visualize these proteins on the surfaces of a series of isogenic strains of E. faecalis. Immunogold labeling, using both 6- and 12-nm colloidal gold, unambiguously demonstrated the expression and distribution of Sec10 and Asc10 on the surface of the E. faecalis cells. On unlabeled E. faecalis cells which expressed either Sec10 or Asc10, the former appeared to be more readily detected. Immunogold labeling of E. faecalis cells expressing both Asc10 and Sec10 clearly demonstrated the abundance and intermixing of both proteins on the cell surface except at septal regions. Sec10 was observed to be distributed over the cell surface. At regions of cell-cell contact, fine strands representing Asc10 were observed directly attaching adjacent cells to one another.
Olmsted
S B
SB
Department of Laboratory Medicine & Pathology, University of Minnesota, Minneapolis 55455-0374.
Erlandsen
S L
SL
Dunny
G M
GM
Wells
C L
CL
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000648:ultrastructure; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000648:ultrastructure; D002462:Cell Membrane / Q000648:ultrastructure; D003227:Conjugation, Genetic; D013293:Enterococcus faecalis / Q000235:genetics* / Q000648:ultrastructure*; D005144:F Factor; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000648:ultrastructure; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods; D016253:Microscopy, Immunoelectron; D010675:Pheromones / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D008565:Membrane Proteins; D010675:Pheromones; C083493:Sec10 protein, Enterococcus faecalis; C071729:Asc10 protein, Enterococcus faecalis