Previous Article | Next Article 
Journal of Bacteriology, April 1999, p. 2459-2464, Vol. 181, No. 8
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The ArgR Regulatory Protein, a Helper to the
Anaerobic Regulator ANR during Transcriptional Activation of the
arcD Promoter in Pseudomonas aeruginosa
Chung-Dar
Lu,1
Harald
Winteler,2
Ahmed
Abdelal,1,* and
Dieter
Haas2
Department of Biology, Georgia State
University, Atlanta, Georgia 30303,1 and
Laboratoire de Biologie Microbienne, Université de
Lausanne, CH-1015 Lausanne, Switzerland2
Received 2 December 1998/Accepted 10 February 1999
 |
ABSTRACT |
Pseudomonas aeruginosa, when deprived of oxygen,
generates ATP from arginine catabolism by enzymes of the arginine
deiminase pathway, encoded by the arcDABC operon. Under
conditions of low oxygen tension, the transcriptional activator ANR
binds to a site centered 41.5 bp upstream of the arcD
transcriptional start. ANR-mediated anaerobic induction was enhanced
two- to threefold by extracellular arginine. This arginine effect
depended, in trans, on the transcriptional regulator ArgR
and, in cis, on an ArgR binding site centered at 
73.5 bp
in the arcD promoter. Binding of purified ArgR protein to
this site was demonstrated by electrophoretic mobility shift assays and
DNase I footprinting. This ArgR recognition site contained a sequence,
5'-TGACGC-3', which deviated in only 1 base from the common
sequence motif 5'-TGTCGC-3' found in other ArgR binding sites of P. aeruginosa. Furthermore, an alignment of all
known ArgR binding sites confirmed that they consist of two directly repeated half-sites. In the absence of ANR, arginine did not induce the
arc operon, suggesting that ArgR alone does not activate
the arcD promoter. According to a model proposed, ArgR
makes physical contact with ANR and thereby facilitates initiation of
arc transcription.
| |
INTRODUCTION |
The arginine deiminase (ADI) pathway
catabolizes L-arginine to L-ornithine, with
concomitant formation of ATP from ADP; three enzymes are involved: ADI,
catabolic ornithine carbamoyltransferase, and carbamate kinase
(17). The ADI pathway can provide Pseudomonas aeruginosa (17, 27) and other bacteria (33)
with energy under anaerobic conditions in the absence of terminal
electron acceptors such as oxygen or nitrate. In P. aeruginosa, the structural genes (arcABC) for the three
enzymes are preceded by a gene (arcD) encoding an
arginine-ornithine antiporter. The four genes are organized as an
operon whose nucleotide sequence has been determined (2, 14,
15). Consistent with its function, the ADI pathway is strongly
induced by oxygen limitation (17). The transcriptional regulator ANR (for anaerobic regulation of arginine catabolism and
nitrate reduction), a homologue of the FNR protein of Escherichia coli, mediates this induction by acting at the 40 region of the arcD promoter (4, 5, 29, 32). Mutants defective
in the arcDABC operon (27) or in ANR
(4) cannot grow anaerobically with arginine as the only
energy source.
In addition to anaerobic control by ANR, exogenous arginine can also
induce the ADI pathway (1, 15, 17). Recent work (21,
22) has characterized a regulatory protein, ArgR, that represses
the arginine biosynthesis carAB and argF genes
encoding carbamoylphosphate synthetase and anabolic ornithine
carbamoyltransferase, respectively. ArgR also mediates induction of the
aruCFGDBE operon, which encodes enzymes of the arginine
succinyltransferase pathway (8, 22). This pathway is
considered to be the major route for arginine catabolism in P. aeruginosa under aerobic conditions (6, 11) and
provides the cell with carbon, nitrogen, and energy. Furthermore, ArgR
was shown to positively control the aotJQMOP-argR operon,
which encodes a system for the uptake of arginine and ornithine as well
as ArgR itself (19).
The mechanism by which arginine induces the ADI pathway in P. aeruginosa has not been studied previously. We have considered two
hypotheses. When terminal electron acceptors are absent,
arginine
being the only energy sourcestimulates the synthesis of
macromolecules and cell growth. Hence, arginine could have a general
stimulatory effect on enzyme induction, including that of the ADI
pathway. Alternatively or additionally, arginine might specifically
enhance the expression of the arc operon. Our present data
support the second hypothesis and establish that the arginine effect is
essentially mediated by ArgR, which binds to a conserved sequence
located upstream of the ANR binding site in the arcD promoter.
| |
MATERIALS AND METHODS |
Bacterial strains, plasmids, and growth conditions.
P.
aeruginosa strains used include PAO1 (wild type), PAO501
(argR::Gmr) (21), PAO6251
(
arcDABC) (5), and PAO6261
[anr(PvuII)] (31). The plasmids
used are listed in Table 1. P. aeruginosa was grown in minimal medium OS (20)
containing 25 mM L-glutamate and 25 mM
L-arginine, but no
(NH4)2SO4. This medium was
supplemented with carbenicillin at 100 µg/ml to maintain recombinant
plasmids and, when PAO501 was the host strain, with gentamicin at 10 µg/ml. Oxygen limitation was achieved in tightly closed bottles as
previously described (4).
Plasmid constructions.
The regulatory region of the
arcDABC operon carried by pME190 (15) was
subcloned after PCR amplification employing two oligonucleotide primers
designed to generate the inborn HindIII site upstream of
the arc promoter (5) and a novel BamHI
site at the 5' end of the arcD gene (Fig.
1). The amplified fragment was digested with HindIII and BamHI and ligated into pUC19
cleaved with the same enzymes. The resulting plasmid was designated as
pAC100. The recombinant PCR procedure described by Higuchi
(7) was employed to modify the ArgR binding sequence in the
arc regulatory region carried by pME190 and its derivative,
pME3731. The amplified PCR fragments were cloned into
HindIII and BamHI sites of pUC19, and the
resulting plasmids were designated pAC102 and pAC101, respectively.
Modification of the 5'-GCGTCA-3' sequence (positions 79 to
74) (Fig. 1) to 5'-ATACTG-3' in each of these two plasmids was confirmed by nucleotide sequencing. The modified sequence of pAC101
was used to replace the ArgR binding sequence in pME3731 by digestion
of the plasmid with HindIII and EcoRI,
treatment with alkaline phosphatase, and ligation with the purified
HindIII-EcoRI fragment of pAC101; this
produced pME3731-1. Plasmid pME6306 was constructed by deletion of the
internal 413-bp SacII fragment in the arcC gene.
The mutated arcC gene was then used to replace the
arcC+ gene in pME190.
View larger version (21K):
[in this window]
[in a new window]
|
FIG. 1.
(a) Schematic drawing of the genetic organization of the
arc operon in the recombinant plasmids used. pME190 carries
the entire wild-type operon, and pME6306 carries a large deletion in
arcC, as described in Table 1. (b) Nucleotide sequence of
the arc regulatory region in pME190 and its derivatives. The
operator sites for ArgR and ANR are indicated by rectangular and oval
boxes, respectively. The HindIII site upstream of the
arc promoter occurs naturally (5). The
BamHI site at the 3' end was introduced by PCR to construct
pAC100 from pME190. Conserved nucleotides are shown in boldface.
|
|
For plasmid transformation into
E. coli DH5
(Bethesda
Research Laboratory) and
P. aeruginosa strains, the method
described
by Chung et al. (
3) for one-step preparation of
competent cells
was
followed.
Enzyme assays.
ADI was assayed by measurements of citrulline
production as described previously (15). Specific activities
are expressed as units (micromoles of citrulline formed per hour) per
milligram of protein. 
-Galactosidase activity was determined by the
Miller method (18).
DNA footprinting and gel retardation experiments.
A DNA
fragment carrying the regulatory region of the arc operon
was obtained from pAC100 by digestion with HindIII and
BamHI endonucleases and was radioactively labeled with DNA
polymerase Klenow fragment, with either [-32P]dATP at
the HindIII site for the bottom strand or with
[-32P]dGTP at the BamHI site for the top strand.
DNase I footprinting was carried out with homogeneous ArgR protein as
previously described (
22). The reaction mixture (200
µl)
contained 2.5 × 10
10 M operator DNA, 2.5 to 20 nM
ArgR protein, 50 mM Tris-HCl (pH
7.5), 50 mM KCl, 10 mM
MgCl
2, 1 mM dithiothreitol, 1 µg of sheared
salmon sperm
DNA, and 10 µg of bovine serum albumin. After incubation
for 30 min
at 25°C, pancreatic DNase I (0.2 µg; Boehringer) was
added. The
digestion was allowed to proceed for 2 min and then
terminated by the
addition of 20 µl of 3 M sodium acetate, 10
µg of yeast tRNA, and
600 µl of ethanol. After precipitation with
ethanol at
70°C, the
pellet was dissolved in 20 µl of formamide-dye
mixture, and the
reaction products were analyzed on a 6% denaturing
polyacrylamide
sequencing gel against a G sequencing ladder (
16).
For gel retardation experiments, the radioactively labeled DNA probe
(5 × 10
12 M) was allowed to interact with different
concentrations of ArgR
in 20 µl of 20 mM Tris-HCl (pH 7.6), 50 mM
KCl, 1 mM EDTA, 5%
(vol/vol) glycerol, and 50 µg of bovine serum
albumin per ml.
The reaction mixtures were allowed to equilibrate for
20 min at
25°C, the reaction was terminated by the addition of an
excess
of cold DNA probe (10
10 M), and then the mixtures
were applied to a 5% polyacrylamide
gel while the gel was running
(
13). The apparent equilibrium
binding constant, defined as
the protein concentration required
for half-maximal binding, was
determined from a plot of the percentage
of DNA bound versus the
protein concentration as previously described
(
13).
| |
RESULTS |
Induction by arginine can occur in the absence of ATP formation
from arginine.
Strain PAO6251, in which the entire
arcDABC operon is deleted, cannot generate ATP from
arginine, unless this mutant is complemented with a plasmid carrying
the intact arc operon (5), such as the IncQ
recombinant plasmid pME190 (Fig. 1). The effect of exogenous arginine
on induction of the arc operon was examined by measurements of ADI, the arcA product, in strain PAO6251 carrying various
plasmids and grown under oxygen limitation and in the absence or
presence of arginine. The results (Table
2) show that exogenous arginine induces
ADI twofold in cells harboring pME190, which carries the intact
arc operon. A similar level of induction is observed in cells harboring pME6306, a derivative of pME190 containing a large deletion in the arcC gene (Fig. 1). This deletion
blocks the function of the ATP-regenerating enzyme, carbamate kinase.
Furthermore, a translational arcD'-'lacZ fusion on
pME336 (5) was induced approximately threefold by arginine.
These results indicate that induction by extracellular arginine is not
merely due to an extra supply of ATP produced from arginine. Therefore,
we tested the hypothesis that arginine could directly affect the
expression of the arc operon.
ArgR protein and an ArgR binding motif in the arc
promoter are required for arginine induction.
Centered at 73.5
nucleotides from the transcription start (5), the
arc promoter contains a sequence motif (Fig. 1) resembling the ArgR binding sites in the aotJ, aruC,
carA, and argF promoters (Fig.
2). The ArgR binding site consists of two
half-sites in a direct repeat arrangement, with the consensus sequence
of 5'-TGTCGCN8AA-3' (22). The most-conserved
nucleotides, 5'-GCGTCA-3', which are located on the
complementary strand at positions 79 to 74 in the arc
promoter, were mutated to 5'-ATACTG-3' (Fig. 1). To
introduce this mutation into the arc promoter, we used a
plasmid (pME3731) containing an artificial EcoRI site in the
30 region of the arc promoter (Fig. 1). We have shown
before that the introduced EcoRI site does not interfere
with anaerobic, ANR-dependent induction of the arc operon
(5). The plasmid containing the mutated ArgR binding motif
was designated pME3731-1.
View larger version (18K):
[in this window]
[in a new window]
|
FIG. 2.
Sequence alignment of ArgR binding sites. The sequences
were obtained from the results of DNase I footprintings and aligned by
using the Clustal W program (25). The first and second
halves of the binding sites (I and II) are depicted by arrows. The
consensus sequence was deduced from the second half-sites, which are
more conserved (19, 21, 22). Nucleotides identical to those
of the consensus site are shaded.  70-RNP, RNA polymerase
holoenzyme; 35 and 10, positions of the promoter recognized by
70. The sequences shown for arcD and
argF are for the complementary strands and are in the
opposite orientation from the direction of transcription.
|
|
ADI activity in the wild-type strain PAO1 and its
argR and
anr derivatives was measured. The results (Table
3) clearly establish
that induction by
exogenous arginine during oxygen limitation
is abolished in either
derivative. Similarly, in PAO1 harboring
pME3731, ADI was inducible in
response to arginine, but to a lower
extent, likely reflecting the
titration of one or both activators
under multicopy conditions (The
IncQ vector used has 20 to 40
copies in
P. aeruginosa)
(
12). In contrast, no arginine induction
was detected in
strain PAO1/pME3731-1 (Table
3), in which the
ArgR binding site was
modified. In the
argR mutant PAO501, both
pME3731 and
pME3731-1 were not induced by arginine (Table
3).
In the
anr
mutant PAO6261 carrying pME3731 or pME3731-1, ADI expression
was low
and was not influenced by extracellular arginine (Table
3). On the one
hand, these data confirm that ANR is the major
transcriptional
regulator of the
arc operon (
5,
32). On the
other
hand, the data suggest that the presence of ANR is necessary
for ArgR
to be an activator of the
arc operon.
ArgR binds to the 70 region of the arc promoter in
vitro.
In order to test whether the ArgR binding motif in the
arc promoter is a functional ArgR binding site, we carried
out gel retardation experiments with homogeneous ArgR protein and a
232-bp HindIII-BamHI fragment carrying the
control region of the arc operon (Fig. 1). The ArgR protein
was found to bind specifically to this arcD regulatory
region (Fig. 3), with an apparent
dissociation constant of 4 × 1011 M as determined
from a plot of the percentage of bound DNA against the concentration of
ArgR (data not shown). When the arc control region
containing the mutated ArgR binding site (see above) was used as a
control, no ArgR-dependent band shift was observed (Fig. 3).
View larger version (22K):
[in this window]
[in a new window]
|
FIG. 3.
Gel retardation experiments. The radioactive
32P-labeled arc operator DNA fragments were
incubated with various ArgR concentrations in the absence of
L-arginine. Lanes 1 to 5 contained the wild-type promoter
fragment (from pAC100) and ArgR at 0, 25, 50, 100, and 200 pM,
respectively. Lanes 6 to 10 contained the promoter fragment mutated in
the ArgR binding site (from pAC102) and ArgR at 0, 25, 50, 100, and 200 pM, respectively.
|
|
DNase I footprinting analysis was used to define the extent of the ArgR
binding site. Binding of ArgR protected a 45-bp region
against nuclease
digestion on both strands (Fig.
4). This
region
was found at the predicted site immediately upstream of the ANR
box (Fig.
1).
View larger version (51K):
[in this window]
[in a new window]
|
FIG. 4.
DNase I footprintings. The DNA fragment was labeled with
[32P]dATP at the 3' end of the HindIII
site on the bottom strand of the arc operator (Fig. 1).
Lanes: 1 and 2, DNase I digestion in the absence of ArgR and in the
presence of 20 nM ArgR, respectively; 3 and 4, 'G' Maxam-Gilbert
sequencing ladder (16); 5 and 6, analogous to lanes 2 and 1, but the DNA fragment was labeled with [32P]dGTP at the 3'
end of the BamHI site on the top strand. The nucleotide
sequence of the ArgR-protected region is shown below the sequencing
ladder, with italicized characters corresponding to the most conserved
bases of the ArgR binding sites,
TGTCGCN8AA.
|
|
| |
DISCUSSION |
The major signal governing the expression of the
arcDABC operon in P. aeruginosa is oxygen
limitation (17). Such conditions allow the ANR protein to
activate transcription (4, 32) by binding to the 40 region
of the arc promoter (5, 29). However, it has been
clear from the early studies of Mercenier et al. (17) and
Abdelal et al. (1) that exogenous arginine can also act as
an inducing signal, although the induction factor is significantly smaller than that brought about by lowering the oxygen tension.
Several lines of evidence reported here show that arginine induction of
the arc operon is mediated by ArgR. (i) Induction by
exogenous arginine is abolished in a derivative of PAO1 in which
argR was inactivated by gene replacement, by using a
gentamicin cassette. (ii) Gel retardation experiments (Fig. 3) show
that ArgR binds specifically to a DNA fragment carrying the control region of the arc operon and that this binding is abolished
when conserved bases in the ArgR binding site are modified. (iii) When the arc promoter contains the modified ArgR binding site,
ADI is no longer induced by exogenous arginine (Table 3).
DNase I footprinting experiments (Fig. 4) show that ArgR protects a
segment of 45 bases located upstream of the arc promoter. The identified binding site was compared with the ArgR binding sites
for the argF, car, aru, and
aot promoters (Fig. 2). This alignment shows that a
conserved sequence (TGACGC) in the ArgR binding site of the
arc operon differs by only 1 base from the consensus
sequence, TGTCGC, which was shown by premethylation and
depurination experiments (21) to be important for ArgR
binding. Apparently, the nonconserved base (A) found in the
arc promoter does not prevent ArgR from binding.
In the arc promoter, the ArgR and ANR binding sites are
adjacent, suggesting that the ArgR and ANR proteins may physically interact with each other during activation of transcription.
Interestingly, the ArgR binding sequence resembling the consensus
sequence is on the bottom strand of the arc regulatory
region. This arrangement differs from that found for other
arginine-inducible operons (aot and aru)
(22), but it occurs in the argF promoter, which
is repressible by ArgR (Fig. 2). It will be interesting to determine if
this spatial arrangement is a prerequisite for ArgR and ANR interactions. In the model presented in Fig.
5, ANR is assumed to interact with RNA
polymerase in exactly the same way as FNR does with RNA polymerase of
E. coli (23). Considering the high degree of
sequence conservation between FNR and ANR (28, 29, 32), this
assumption appears justified. In oxygen-limited cells, ANR can activate
transcription without arginine and ArgR (15, 17, 29) (Table
2). Arginine and ArgR boost the ANR-dependent induction (Table 3). In
well-aerated cells (17) or in an ANR-negative mutant (Table
3), arginine does not induce the arc operon, suggesting that
ArgR alone cannot activate arc transcription. This situation is reminiscent of nitrate respiration in E. coli in which
the anaerobic activator FNR is assisted by the nitrate- and
nitrite-responsive response regulators NarL and NarP at the
nir and nrf promoters (24, 26).
View larger version (11K):
[in this window]
[in a new window]
|
FIG. 5.
Model depicting the interaction between ArgR and ANR at
the arc promoter of P. aeruginosa. Given the
overlapping specificity of FNR and ANR and the functional conservation
of the two regulators (28, 29, 32), this model is based on
the structural data elaborated for FNR and RNA polymerase of E. coli (23). The 70, , and '
subunits of the RNA polymerase were labeled accordingly. CTD and
NTD represent the carboxy-terminal domain and the amino-terminal
domain of the subunits, respectively. The possible contact between
ANR and the 70 subunit is represented by a solid
square.
|
|
Recently, Maghnouj et al. (15a) reported the cloning and
characterization of the arc operon from Bacillus
licheniformis. While anaerobiosis is a prerequisite for arginine
induction of the arc operon of P. aeruginosa, as
outlined above, both anaerobiosis and arginine, as well as a functional
ArgR, were required for induction of the arc operon in
B. licheniformis (15a). ArgR of B. licheniformis was shown to bind a sequence homologous to the Arg
boxes that characterize operator sites for the arginine repressors from
enteric bacteria and B. subtilis. A nucleotide sequence
feature similar to the E. coli Crp and B. subtilis Fnr consensus sequences was found between the ArgR
binding site and the promoter, suggesting that a second regulatory
protein from the Crp or Fnr family might function as the anaerobic
activator of this system. It should be noted that ArgR of B. licheniformis is a hexamer of a 17-kDa polypeptide, as is the case
for the arginine regulatory proteins from enteric bacteria and B. subtilis; these proteins also contain highly conserved domains
(15a). In contrast, the ArgR of P. aeruginosa is
a dimer of 37 kDa that belongs to the AraC-XylS family and does not
exhibit sequence similarity to the regulatory proteins of enteric
bacteria or B. subtilis (21, 22).
In vitro, the presence of arginine did not enhance binding of ArgR to
the arc promoter from P. aeruginosa (data not
shown). Previous work (19, 21, 22) has shown that arginine
had no effect on the affinity of ArgR for the car,
argF, aru, and aot promoters. Two
interpretations can be offered. Arginine might facilitate the
interaction between ArgR and the ANR-RNA polymerase complex, without
influencing the binding of ArgR to its operator sequence.
Alternatively, intracellular arginine might not be the relevant signal.
Instead, extracellular arginine might be sensed by a sensor protein,
which, via a response regulator, would activate the expression of the
aot-argR operon. This hypothesis is supported by the recent
identification of the ArgSU two-component system in P. aeruginosa. Mutational inactivation of the sensor ArgS results in
an arginine-utilization-negative phenotype, presumably because of loss
of ArgR synthesis (9), and in an argU-negative
background, an aot-lacZ fusion is not expressed
(10).
| |
ACKNOWLEDGMENTS |
We thank Marianne Gamper for discussion. Carbenicillin was a gift
from SmithKline Beecham.
Support from EU project BIO4-CT96-0119 and NIH research grant GM47926
is gratefully acknowledged.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Dean's Office,
Georgia State University, P.O. Box 4038, Atlanta, GA 30302-4038. Phone: (404) 651-1410. Fax: (404) 651-4739. E-mail:
aabdelal{at}gsu.edu.
| |
REFERENCES |
| 1.
|
Abdelal, A. T.,
W. F. Bibb, and O. Nainan.
1982.
Carbamate kinase from Pseudomonas aeruginosa: purification, characterization, physiological role, and regulation.
J. Bacteriol.
151:1411-1419[Abstract/Free Full Text].
|
| 2.
|
Baur, H.,
E. Luthi,
V. Stalon,
A. Mercenier, and D. Haas.
1989.
Sequence analysis and expression of the arginine-deiminase and carbamate-kinase genes of Pseudomonas aeruginosa.
Eur. J. Biochem.
179:53-60[Medline].
|
| 3.
|
Chung, C. T.,
S. L. Niemela, and R. H. Miller.
1989.
One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution.
Proc. Natl. Acad. Sci. USA
86:2172-2175[Abstract/Free Full Text].
|
| 4.
|
Galimand, M.,
M. Gamper,
A. Zimmermann, and D. Haas.
1991.
Positive FNR-like control of anaerobic arginine degradation and nitrate respiration in Pseudomonas aeruginosa.
J. Bacteriol.
173:1598-1606[Abstract/Free Full Text].
|
| 5.
|
Gamper, M.,
A. Zimmermann, and D. Haas.
1991.
Anaerobic regulation of transcription initiation in the arcDABC operon of Pseudomonas aeruginosa.
J. Bacteriol.
173:4742-4750[Abstract/Free Full Text].
|
| 6.
|
Haas, D.,
H. Winteler,
V. T. Nguyen,
C. Tricot, and V. Stalon.
1997.
Arginine catabolism in Pseudomonas aeruginosa: key regulatory features of the arginine deiminase pathway, p. 389-399.
In
P. P. De Deyn, B. Marescau, I. A. Qureshi, and A. Mori (ed.), Guanido compounds in biology and medicine, vol. 2. John Libbey, London, United Kingdom.
|
| 7.
|
Higuchi, R.
1990.
Recombinant PCR, p. 177-183.
In
M. A. Innis, D. H. Gelfand, J. J. Sninsky, and T. J. White (ed.), PCR protocols. Academic Press, New York, N.Y.
|
| 8.
|
Itoh, Y.
1997.
Cloning and characterization of the aru genes encoding enzymes of the catabolic arginine succinyltransferase pathway in Pseudomonas aeruginosa.
J. Bacteriol.
179:7280-7290[Abstract/Free Full Text].
|
| 9.
|
Itoh, Y.,
T. Nishijyo,
S.-M. Park, and D. Haas.
1998.
Novel regulatory genes involved in arginine and ornithine catabolism in Pseudomonas aeruginosa, abstr. 39.
In
Proceedings of the XVIth International Arginine/Pyrimidine Conference. Leeds, United Kingdom..
|
| 10.
| Itoh, Y. Personal communication.
|
| 11.
|
Jann, A.,
V. Stalon,
C. Vander Wauven,
T. Leisinger, and D. Haas.
1986.
N2-succinylated intermediates in an arginine catabolic pathway of Pseudomonas aeruginosa.
Proc. Natl. Acad. Sci. USA
83:4937-4941[Abstract/Free Full Text].
|
| 12.
|
Jeenes, D. J.,
L. Soldati,
H. Baur,
J. M. Watson,
A. Mercenier,
C. Reimmann,
T. Leisinger, and D. Haas.
1986.
Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host.
Mol. Gen. Genet.
203:421-429[Medline].
|
| 13.
|
Lu, C. D.,
J. E. Houghton, and A. T. Abdelal.
1992.
Characterization of the arginine repressor from Salmonella typhimurium and its interactions with the carAB operator.
J. Mol. Biol.
225:11-24[Medline].
|
| 14.
|
Lüthi, E.,
H. Baur,
M. Gamper,
F. Brunner,
D. Villeval,
A. Mercenier, and D. Haas.
1990.
The arc operon for anaerobic arginine catabolism in Pseudomonas aeruginosa contains an additional gene, arcD, encoding a membrane protein.
Gene
87:37-43[Medline].
|
| 15.
|
Lüthi, E.,
A. Mercenier, and D. Haas.
1986.
The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes.
J. Gen. Microbiol.
132:2667-2675[Medline].
|
| 15a.
|
Maghnouj, A.,
T. F. de Sousa Cabral,
V. Stalon, and C. Vander Wauven.
1998.
The arcABDC gene cluster, encoding the arginine deiminase pathway of Bacillus licheniformis, and its activation by the arginine repressor ArgR.
J. Bacteriol.
180:6468-6475[Abstract/Free Full Text].
|
| 16.
|
Maxam, A. M., and W. Gilbert.
1980.
Sequencing end-labeled DNA with base-specific cleavages.
Methods Enzymol.
65:499-560[Medline].
|
| 17.
|
Mercenier, A.,
J.-P. Simon,
C. Vander Wauven,
D. Haas, and V. Stalon.
1980.
Regulation of enzyme synthesis in the arginine deiminase pathway of Pseudomonas aeruginosa.
J. Bacteriol.
144:159-163[Abstract/Free Full Text].
|
| 18.
|
Miller, J. H.
1972.
Experiments in molecular genetics.
Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
|
| 19.
|
Nishijyo, T.,
S.-M. Park,
C.-D. Lu,
Y. Itoh, and A. T. Abdelal.
1998.
Molecular characterization and regulation of an operon encoding a system for transport of arginine and ornithine and the ArgR regulatory protein in Pseudomonas aeruginosa.
J. Bacteriol.
180:5559-5566[Abstract/Free Full Text].
|
| 20.
|
Ornston, L., and R. Y. Stanier.
1966.
The conversion of catechol and protocatechuate to -ketoadipate by Pseudomonas putida.
J. Biol. Chem.
241:3776-3786[Abstract/Free Full Text].
|
| 21.
|
Park, S.-M.,
C.-D. Lu, and A. T. Abdelal.
1997.
Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1.
J. Bacteriol.
179:5300-5308[Abstract/Free Full Text].
|
| 22.
|
Park, S.-M.,
C.-D. Lu, and A. T. Abdelal.
1997.
Purification and characterization of an arginine regulatory protein, ArgR, from Pseudomonas aeruginosa and its interactions with the control regions for the car, argF, and aru operons.
J. Bacteriol.
179:5309-5317[Abstract/Free Full Text].
|
| 23.
|
Rhodius, V. A., and S. J. W. Busby.
1998.
Positive activation of gene expression.
Curr. Opin. Microbiol.
1:152-159.
[Medline] |
| 24.
|
Stewart, V., and R. S. Rabin.
1995.
Dual sensors and dual response regulators interact to control nitrate- and nitrite-responsive gene expression in Escherichia coli, p. 233-252.
In
J. A. Hoch, and T. Silhavy (ed.), Two-component signal transduction. ASM Press, Washington, D.C.
|
| 25.
|
Thompson, J. D.,
D. G. Higgins, and T. J. Gilson.
1994.
Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
Nucleic Acids Res.
22:4674-4680.
|
| 26.
|
Tyson, K. L.,
J. A. Cole, and J. W. Busby.
1994.
Nitrite and nitrate regulation at the promoters of two Escherichia coli operons encoding nitrite reductase: identification of common target heptamers for both NarP- and NarL-dependent regulation.
Mol. Microbiol.
13:1045-1055[Medline].
|
| 27.
|
Vander Wauven, C.,
A. Piérard,
M. Kley-Raymann, and D. Haas.
1984.
Pseudomonas aeruginosa mutants affected in anaerobic growth on arginine: evidence for a four-gene cluster encoding the arginine deiminase pathway.
J. Bacteriol.
160:928-934[Abstract/Free Full Text].
|
| 28.
|
Williams, S. M.,
N. J. Savery,
S. J. W. Busby, and H. J. Wing.
1997.
Transcription activation at class I FNR-dependent promoters: identification of the activating surface of FNR and the corresponding contact site in the C-terminal domain of the RNA polymerase subunit.
Nucleic Acids Res.
25:4028-4034[Abstract/Free Full Text].
|
| 29.
|
Winteler, H., and D. Haas.
1996.
The homologous regulators ANR of Pseudomonas aeruginosa and FNR of Escherichia coli have overlapping but distinct specificities for anaerobically inducible promoters.
Microbiology
142:685-693[Abstract/Free Full Text].
|
| 30.
|
Yanisch-Perron, C.,
J. Vieira, and J. Messing.
1985.
Improved M13 phage cloning vectors and host strains: nucleotide sequences of M13mp18 and pUC19 vectors.
Gene
33:103-119[Medline].
|
| 31.
|
Ye, R. W.,
D. Haas,
J.-O. Ka,
V. Krishnapillai,
A. Zimmermann,
C. Baird, and J. M. Tiedje.
1995.
Anaerobic activation of the entire denitrification pathway in Pseudomonas aeruginosa requires Anr, an analog of Fnr.
J. Bacteriol.
177:3606-3609[Abstract/Free Full Text].
|
| 32.
|
Zimmermann, A.,
C. Reimmann,
M. Galimand, and D. Haas.
1991.
Anaerobic growth and cyanide synthesis of Pseudomonas aeruginosa depend on anr, a regulatory gene homologous with fnr of Escherichia coli.
Mol. Microbiol.
5:1483-1490[Medline].
|
| 33.
|
Zúñiga, M.,
M. Champomier-Verges,
M. Zagorec, and G. Pérez-Martinez.
1998.
Structural and functional analysis of the gene cluster encoding the enzymes of the arginine deiminase pathway of Lactobacillus sake.
J. Bacteriol.
180:4154-4159[Abstract/Free Full Text].
|
Journal of Bacteriology, April 1999, p. 2459-2464, Vol. 181, No. 8
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Benkert, B., Quack, N., Schreiber, K., Jaensch, L., Jahn, D., Schobert, M.
(2008). Nitrate-responsive NarX-NarL represses arginine-mediated induction of the Pseudomonas aeruginosa arginine fermentation arcDABC operon. Microbiology
154: 3053-3060
[Abstract]
[Full Text]
-
Liu, Y., Dong, Y., Chen, Y.-Y. M., Burne, R. A.
(2008). Environmental and Growth Phase Regulation of the Streptococcus gordonii Arginine Deiminase Genes. Appl. Environ. Microbiol.
74: 5023-5030
[Abstract]
[Full Text]
-
Platt, M. D., Schurr, M. J., Sauer, K., Vazquez, G., Kukavica-Ibrulj, I., Potvin, E., Levesque, R. C., Fedynak, A., Brinkman, F. S. L., Schurr, J., Hwang, S.-H., Lau, G. W., Limbach, P. A., Rowe, J. J., Lieberman, M. A., Barraud, N., Webb, J., Kjelleberg, S., Hunt, D. F., Hassett, D. J.
(2008). Proteomic, Microarray, and Signature-Tagged Mutagenesis Analyses of Anaerobic Pseudomonas aeruginosa at pH 6.5, Likely Representing Chronic, Late-Stage Cystic Fibrosis Airway Conditions. J. Bacteriol.
190: 2739-2758
[Abstract]
[Full Text]
-
Seggewiss, J., Becker, K., Kotte, O., Eisenacher, M., Yazdi, M. R. K., Fischer, A., McNamara, P., Al Laham, N., Proctor, R., Peters, G., Heinemann, M., von Eiff, C.
(2006). Reporter Metabolite Analysis of Transcriptional Profiles of a Staphylococcus aureus Strain with Normal Phenotype and Its Isogenic hemB Mutant Displaying the Small-Colony-Variant Phenotype. J. Bacteriol.
188: 7765-7777
[Abstract]
[Full Text]
-
Schaumburg, C. S., Tan, M.
(2006). Arginine-Dependent Gene Regulation via the ArgR Repressor Is Species Specific in Chlamydia. J. Bacteriol.
188: 919-927
[Abstract]
[Full Text]
-
Zeng, L., Dong, Y., Burne, R. A.
(2006). Characterization of cis-Acting Sites Controlling Arginine Deiminase Gene Expression in Streptococcus gordonii. J. Bacteriol.
188: 941-949
[Abstract]
[Full Text]
-
Gruening, P., Fulde, M., Valentin-Weigand, P., Goethe, R.
(2006). Structure, Regulation, and Putative Function of the Arginine Deiminase System of Streptococcus suis. J. Bacteriol.
188: 361-369
[Abstract]
[Full Text]
-
Hashim, S., Kwon, D.-H., Abdelal, A., Lu, C.-D.
(2004). The Arginine Regulatory Protein Mediates Repression by Arginine of the Operons Encoding Glutamate Synthase and Anabolic Glutamate Dehydrogenase in Pseudomonas aeruginosa. J. Bacteriol.
186: 3848-3854
[Abstract]
[Full Text]
-
Lu, C.-D., Yang, Z., Li, W.
(2004). Transcriptome Analysis of the ArgR Regulon in Pseudomonas aeruginosa. J. Bacteriol.
186: 3855-3861
[Abstract]
[Full Text]
-
Dong, Y., Chen, Y.-Y. M., Burne, R. A.
(2004). Control of Expression of the Arginine Deiminase Operon of Streptococcus gordonii by CcpA and Flp. J. Bacteriol.
186: 2511-2514
[Abstract]
[Full Text]
-
Galkin, A., Kulakova, L., Sarikaya, E., Lim, K., Howard, A., Herzberg, O.
(2004). Structural Insight into Arginine Degradation by Arginine Deiminase, an Antibacterial and Parasite Drug Target. J. Biol. Chem.
279: 14001-14008
[Abstract]
[Full Text]
-
Zuniga, M., Miralles, M. d. C., Perez-Martinez, G.
(2002). The Product of arcR, the Sixth Gene of the arc Operon of Lactobacillus sakei, Is Essential for Expression of the Arginine Deiminase Pathway. Appl. Environ. Microbiol.
68: 6051-6058
[Abstract]
[Full Text]
-
Barcelona-Andres, B., Marina, A., Rubio, V.
(2002). Gene Structure, Organization, Expression, and Potential Regulatory Mechanisms of Arginine Catabolism in Enterococcus faecalis. J. Bacteriol.
184: 6289-6300
[Abstract]
[Full Text]
-
Dong, Y., Chen, Y.-Y. M., Snyder, J. A., Burne, R. A.
(2002). Isolation and Molecular Analysis of the Gene Cluster for the Arginine Deiminase System from Streptococcus gordonii DL1. Appl. Environ. Microbiol.
68: 5549-5553
[Abstract]
[Full Text]
-
Lu, C.-D., Itoh, Y., Nakada, Y., Jiang, Y.
(2002). Functional Analysis and Regulation of the Divergent spuABCDEFGH-spuI Operons for Polyamine Uptake and Utilization in Pseudomonas aeruginosa PAO1. J. Bacteriol.
184: 3765-3773
[Abstract]
[Full Text]
-
Nakada, Y., Itoh, Y.
(2002). Characterization and Regulation of the gbuA Gene, Encoding Guanidinobutyrase in the Arginine Dehydrogenase Pathway of Pseudomonas aeruginosa PAO1. J. Bacteriol.
184: 3377-3384
[Abstract]
[Full Text]
-
Sauer, K., Camper, A. K., Ehrlich, G. D., Costerton, J. W., Davies, D. G.
(2002). Pseudomonas aeruginosa Displays Multiple Phenotypes during Development as a Biofilm. J. Bacteriol.
184: 1140-1154
[Abstract]
[Full Text]
-
Nakada, Y., Jiang, Y., Nishijyo, T., Itoh, Y., Lu, C.-D.
(2001). Molecular Characterization and Regulation of the aguBA Operon, Responsible for Agmatine Utilization in Pseudomonas aeruginosa PAO1. J. Bacteriol.
183: 6517-6524
[Abstract]
[Full Text]
-
Lu, C.-D., Abdelal, A. T.
(2001). The gdhB Gene of Pseudomonas aeruginosa Encodes an Arginine-Inducible NAD+-Dependent Glutamate Dehydrogenase Which Is Subject to Allosteric Regulation. J. Bacteriol.
183: 490-499
[Abstract]
[Full Text]
-
Pessi, G., Haas, D.
(2000). Transcriptional Control of the Hydrogen Cyanide Biosynthetic Genes hcnABC by the Anaerobic Regulator ANR and the Quorum-Sensing Regulators LasR and RhlR in Pseudomonas aeruginosa. J. Bacteriol.
182: 6940-6949
[Abstract]
[Full Text]
-
Højberg, O., Schnider, U., Winteler, H. V., Sørensen, J., Haas, D.
(1999). Oxygen-Sensing Reporter Strain of Pseudomonas fluorescens for Monitoring the Distribution of Low-Oxygen Habitats in Soil. Appl. Environ. Microbiol.
65: 4085-4093
[Abstract]
[Full Text]
-
Ochs, M. M., Lu, C.-D., Hancock, R. E. W., Abdelal, A. T.
(1999). Amino Acid-Mediated Induction of the Basic Amino Acid-Specific Outer Membrane Porin OprD from Pseudomonas aeruginosa. J. Bacteriol.
181: 5426-5432
[Abstract]
[Full Text]
Top
Abstract
Introduction
