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Journal of Bacteriology, April 1999, p. 2217-2224, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Genes Coding for Phosphotransacetylase and Acetate
Kinase in Sinorhizobium meliloti Are in an Operon That Is
Inducible by Phosphate Stress and Controlled by PhoB
Michael L.
Summers,
Michael
C.
Denton, and
Timothy R.
McDermott*
Department of Land Resources and
Environmental Sciences, Montana State University, Bozeman, Montana
59717
Received 27 October 1998/Accepted 27 January 1999
Recent work in this laboratory has shown that the gene coding for
acetate kinase (ackA) in Sinorhizobium meliloti
is up-regulated in response to phosphate limitation. Characterization
of the region surrounding ackA revealed that it is adjacent
to pta, which codes for phosphotransacetylase, and that
these two genes are part of an operon composed of at least two
additional genes in the following order: an open reading frame
(orfA), pta, ackA, and the partial sequence of a gene with an inferred peptide that has a high degree of
homology to enoyl-ACP reductase (fabI). Experiments
combining enzyme assays, a chromosomal
lacZ::ackA transcriptional fusion, complementation analysis with cosmid subclones, and the creation of
mutations in pta and ackA all indicated that
the orfA-pta-ackA-fabI genes are cotranscribed in response
to phosphate starvation. Primer extension was used to map the position
of the phosphate starvation-inducible transcriptional start sites
upstream of orfA. The start sites were found to be preceded
by a sequence having similarity to PHO boxes from other
phosphate-regulated genes in S. meliloti and to the
consensus PHO box in Escherichia coli. Introduction of a
phoB mutation in the wild-type strain eliminated elevated
levels of acetate kinase and phosphotransacetylase activities in
response to phosphate limitation and also eliminated the phosphate
stress-induced up-regulation of the
ackA::lacZ fusion. Mutations in
either ackA alone or both pta and
ackA did not affect the nodulation or nitrogen fixation
phenotype of S. meliloti.
*
Corresponding author. Mailing address: Department of
Land Resources and Environmental Sciences, Montana State University, Bozeman, MT 59717. Phone: (406) 994-2190. Fax: (406) 994-3933. E-mail:
timmcder{at}gemini.oscs.montana.edu.

Present address: Department of Biology, California State
University

Northridge, Northridge, CA 91330-8303.
Journal of Bacteriology, April 1999, p. 2217-2224, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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