Two novel flagellar components and H-NS are involved in the motor function of Escherichia coli

Abstract

A mutation in H-NS results in non-flagellation of Escherichia coli due to a reduced expression of the flhDC master operon. We found that the hns-negative strain restored its flagellation in the presence of flhDC, although the resulting strain was still non-motile. Since the intracelluar levels of motor components MotA, MotB, and FliG in the Delta hns strain were unaltered, the non-motility indicates that H-NS affects flagellar function as well as biogenesis. We obtained an insertion in ycgR, a putative gene encoding a protein of 244 amino acid residues, which suppresses the motility defect of hns-deficient cells. The abnormally low swimming speed of hns mutant cells was fully restored by an insertion in ycgR, as assessed with computer-assisted motion analysis. A similar suppressor phenotype was observed with a multicopy expression of yhjH, a putative gene encoding a polypeptide of 256 amino acid residues. Since the flagella of most hns-deficient cells were not rotating, except a few with reduced speed, the suppression appears to increase the number of rotating flagella as observed with tethered bacteria. The ycgR and yhjH genes contain the consensus sequence found among the class III promoters of the flagellar regulon, and their expression monitored with a lacZ fusion requires FlhDC. These findings suggest that ycgR and yhjH, together with H-NS, are involved in the motor function and constitute new members of the flagellar regulon. (C) 2000 Academic Press.