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Journal of Bacteriology, August 1999, p. 4761-4767, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The icfG Gene Cluster of
Synechocystis sp. Strain PCC 6803 Encodes an Rsb/Spo-Like
Protein Kinase, Protein Phosphatase, and Two
Phosphoproteins
Liang
Shi,
Kenneth M.
Bischoff, and
Peter J.
Kennelly*
Department of Biochemistry and the Institute
for Genomics, Virginia Polytechnic Institute and State University,
Blacksburg, Virginia 24061-0308
Received 23 March 1999/Accepted 31 May 1999
A set of open reading frames (ORFs) potentially encoding signal
transduction proteins are clustered around icfG, a gene
implicated in the regulation of carbon metabolism, in the genome of
Synechocystis sp. strain PCC 6803. slr1860 is
the ORF for icfG, whose predicted product resembles the
protein phosphatases SpoIIE, RsbU, and RsbX from Bacillus
subtilis. Bracketing slr1860/icfG are (i) ORF
slr1861, whose predicted product resembles the SpoIIAB,
RsbT, and RsbW protein kinases from B. subtilis, and (ii)
ORFs slr1856 and slr1859, whose predicted
products resemble the respective phosphoprotein substrates for the
B. subtilis protein kinases: SpoIIAA, RsbS, and RsbV. In
order to determine whether the protein products encoded by these ORFs
possessed the functional capabilities suggested by sequence
comparisons, each was expressed in Escherichia coli as a
histidine-tagged fusion protein and analyzed for its ability to
participate in protein phosphorylation-dephosphorylation processes in
vitro. It was observed that ORF slr1861 encoded an
ATP-dependent protein kinase capable of phosphorylating Slr1856 and,
albeit with noticeably lower efficiency, Slr1859. Site-directed
mutagenesis suggests that Slr1861 phosphorylated these proteins on
Ser-54 and Ser-57, respectively. Slr1860 exhibited divalent metal
ion-dependent protein-serine phosphatase activity. It catalyzed the
dephosphorylation of Slr1856, but not Slr1859, in vitro.
*
Corresponding author. Mailing address: Department of
Biochemistry and the Institute for Genomics, Virginia Polytechnic
Institute and State University, Blacksburg, VA 24061-0308. Phone: (540) 231-4317. Fax: (540) 231-9070. E-mail: pjkennel{at}vt.edu.
Journal of Bacteriology, August 1999, p. 4761-4767, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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