Journal of Bacteriology, May 2000, p. 2709-2715, Vol. 182, No. 10
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada,1 and Max-Planck-Institut für Molekulare Genetik, Dahlem, D-14195 Berlin, Germany2
Received 1 December 1999/Accepted 24 February 2000
The physical association of bacteria during conjugation mediated by
the IncP
plasmid RP4 was investigated. Escherichia coli mating aggregates prepared on semisolid medium were ultrarapidly frozen
using copper block freezing, followed by freeze substitution, thin
sectioning, and transmission electron microscopy. In matings where the
donor bacteria contained conjugative plasmids, distinctive junctions
were observed between the outer membranes of the aggregates of mating
cells. An electron-dense layer linked the stiffly parallel outer
membranes in the junction zone, but there were no cytoplasmic bridges
nor apparent breaks in the cell walls or membranes. In control
experiments where the donors lacked conjugative plasmids, junctions
were not observed. Previous studies have shown that plasmid RP4 carries
operons for both plasmid DNA processing (Tra1) and mating pair
formation (Tra2). In matings where donor strains carried Tra2 only or
Tra2 plus the pilin-processing protease TraF, junctions were found but
they were shorter and more interrupted than the wild type. If the donor
strain had the pilin gene knocked out (trbC), junctions
were still found. Thus, it appears that the electron-dense layer
between the outer membranes of the conjugating cells is not composed of pilin.
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