The regulator of sigma D (Rsd) was identified as an RNA polymerase sigma(70)-associated protein in stationary-phase Escherichia coli with the inhibitory activity of sigma(70)-dependent transcription in vitro (M, Jishage and A, Ishihama, Proc. Natl. Acad, Sci, USA 95:4953-1958, 1998), Primer extension analysis of rsd mRNA indicated the presence of two promoters, sigma(S)-dependent P1 and sigma(70)-dependent P2 with the gearbox sequence, To get insight into the in vivo role of Rsd, the expression of a reporter gene fused to either the sigma(70)- or as dependent promoter was analyzed in the absence of Rsd or the presence of overexpressed Rsd. In the rsd null mutant, the sigma(70)- and sigma(S)-dependent gene expression was increased or decreased, respectively. On the other hand, the sigma(70)- or sigma(S)-dependent transcription was reduced or enhanced, respectively, after overexpression of Rsd, The repression of the sigma(S)-dependent transcription in the rsd mutant is overcome by increased production of the sigma(S) subunit. Together these observations support the prediction that Rsd is involved in replacement of the RNA polymerase sigma subunit from sigma(70) to sigma(S) during the transition from exponential growth to the stationary phase.