Mini-F encoded proteins: identification of a new 10.5 kilodalton species.

The elements which ensure the maintenance of the F plasmid are located in its f5 EcoRI restriction fragment. This f5 fragment constitutes a mini-F plasmid showing the same stability and copy number control as the entire F plasmid. The proteins expressed in minicells by wild-type or mutated f5 fragments were analysed by pH gradient two-dimensional electrophoresis. We identified seven f5-encoded polypeptides and located their genes on the F map. Among them, H1, an acidic polypeptide of mol. wt. 10.5 K, had not been detected before. It is in fact the most abundant f5-encoded polypeptide identified so far. In addition, we showed that both 10.5-K and 12-K protein bands detected by SDS-polyacrylamide gel electrophoresis are, respectively, composed of two polypeptides, H1 and H2, G1 and G2, of different isoelectric points. Polypeptides H2 and G2, respectively, share common coding sequences with polypeptides H1 and G1. Their possible biological significance is discussed. The sequences coding for polypeptides H1/H2 and G1/G2 are clustered in a 800-bp long region located between the two mini-F origin sites and are proposed to be organized as an operon. The results reported in the accompanying paper point out the importance of polypeptides G1/G2 and H1/H2 in the relationship between the F plasmid and its host.