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Journal of Bacteriology, August 2000, p. 4478-4490, Vol. 182, No. 16
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Phosphate Starvation-Inducible Proteins of
Bacillus subtilis: Proteomics and Transcriptional
Analysis
Haike
Antelmann,
Christian
Scharf, and
Michael
Hecker*
Institut für Mikrobiologie,
Ernst-Moritz-Arndt-Universität, 17487 Greifswald, Germany
Received 2 March 2000/Accepted 17 May 2000
The phosphate starvation response in Bacillus subtilis
was analyzed using two-dimensional (2D) polyacrylamide gel
electrophoresis of cell extracts and supernatants from
phosphate-starved cells. Most of the phosphate starvation-induced
proteins are under the control of
B, the activity of
which is increased by energy depletion. In order to define the proteins
belonging to the Pho regulon, which is regulated by the two-component
regulatory proteins PhoP and PhoR, the 2D protein pattern of the wild
type was compared with those of a sigB mutant and a
phoR mutant. By matrix-assisted laser desorption ionization-time of flight mass spectrometry, two alkaline phosphatases (APases) (PhoA and PhoB), an APase-alkaline phosphodiesterase (PhoD), a
glycerophosphoryl diester phosphodiesterase (GlpQ), and the lipoprotein
YdhF were identified as very strongly induced PhoPR-dependent proteins
secreted into the extracellular medium. In the cytoplasmic fraction,
PstB1, PstB2, and TuaD were identified as already known PhoPR-dependent
proteins, in addition to PhoB, PhoD, and the previously described PstS.
Transcriptional studies of glpQ and ydhF
confirmed the strong PhoPR dependence. Northern hybridization and
primer extension experiments showed that glpQ is
transcribed monocistronically from a
A promoter which is
overlapped by four putative TT(A/T)ACA-like PhoP binding sites.
Furthermore, ydhF might be cotranscribed with phoB initiating from the phoB promoter. Only a
small group of proteins remained phosphate starvation inducible in both
phoR and sigB mutant and did not form a unique
regulation group. Among these, YfhM and YjbC were controlled by
B-dependent and unknown PhoPR-independent mechanisms.
Furthermore, YtxH and YvyD seemed to be induced after phosphate
starvation in the wild type in a
B-dependent manner and
in the sigB mutant probably via
H. YxiE was
induced by phosphate starvation independently of
B and PhoPR.
*
Corresponding author. Mailing address: Institut
für Mikrobiologie, Ernst-Moritz-Arndt-Universität,
17487 Greifswald, Germany. Phone: 49 (3834) 864200. Fax: 49 (3834) 864202. E-mail:
hecker{at}microbio7.biologie.uni-greifswald.de.
Journal of Bacteriology, August 2000, p. 4478-4490, Vol. 182, No. 16
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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