In Pseudomonas aeruginosa, iron modulates gene expression through a cascade
of negative and positive regulatory proteins. The master regulator Fur is
involved in iron-dependent repression of several genes. One of these genes,
pvdS, was predicted to encode a putative sigma factor responsible for the
transcription of a subset of genes of the Fur regulon. PvdS appears to belo
ng to a structurally and functionally distinct subgroup of the extracytopla
smic function family of alternative sigma factors. Members of this subgroup
, also including PbrA from Pseudomonas fluorescens, PfrI and PupI from Pseu
domonas putida, and FecI from Escherichia coli, are controlled by the Fur r
epressor, and they activate transcription of genes for the biosynthesis or
the uptake of siderophores. Evidence is provided that the PvdS protein of P
. aeruginosa is endowed with biochemical properties of eubacterial sigma fa
ctors, as it spontaneously forms 1:1 complexes with the core fraction of RN
A polymerase (RNAP, alpha(2)beta beta' subunits), thereby promoting in vitr
o binding of the PVdS-RNAP holoenzyme to the promoter region of the pvdA ge
ne. These functional features of PvdS are consistent with the presence of s
tructural domains predicted to be involved in core RNAP binding, promoter r
ecognition, and open complex formation. The activity of pyoverdin biosynthe
tic (pvd) promoters was significantly lower in E. coli overexpressing the m
ulticopy pvdS gene than in wild-type P. aeruginosa PAO1 carrying the single
gene copy, and pvd::lacZ transcriptional fusions were silent in both pfrI
(the pvdS homologue) and pfrA (a positive regulator of pseudobactin biosynt
hetic genes) mutants of P. putida WCS358, while they are expressed at PAO1
levels in wild-type WCS358. Moreover, the PvdS-RNAP holoenzyme purified fro
m E. coli lacked the ability to generate in vitro transcripts from the pvdA
promoter. These observations suggest that at least one additional positive
regulator could be required for full activity of the PvdS-dependent transc
ription complex both in vivo and in vitro. This is consistent with the pres
ence of a putative activator binding site (the iron starvation box) at vari
able distance from the transcription initiation sites of promoters controll
ed by the iron starvation sigma factors PvdS, PfrI, and PbrA of fluorescent
pseudomonads.