Abstract

Inactivation of either of the two MetR binding sites centered at bp —143 and —121 relative to the +1 transcription start site results in reduced glyA-lacZ expression in a wild-type strain below the level seen in a metR mutant. This reduced expression is dependent on the side of the DNA helix MetR binds relative to the RNA polymerase binding site. Thus, a single MetR dimer bound to the DNA may play a physiological role in maintaining appropriate glyA gene expression, functioning as a represser under low MetR conditions.

References

[1]
Stauffer
G.V.
(
1996
)
Biosynthesis of serine, glycine and onecarbon units
. In:
Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology
  (
Neidhardt
F.C.
Curtiss
R.
Ingraham
J.L.
Edmund
C.C.L.
Low
K.B.
Magasanik
B.
Reznikoff
W.S.
Riley
M.
Schaechter
M.
Umbarger
H.E.
, Eds.), pp.
506
513
.
American Society for Microbiology
,
Washington, DC
.
[2]
Schell
M.A.
(
1993
)
Molecular biology of the LysR family of transcriptional regulators
.
Annu. Rev. Microbiol.
 
47
,
597
626
.
[3]
Greene
R.C.
(
1996
)
Biosynthesis of methionine
In:
Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology
  (
Neidhardt
F.C.
Curtiss
R.
Ingraham
J.L.
Edmund
C.C.L.
Low
K.B.
Magasanik
B.
Reznikoff
W.S.
Riley
M.
Schaechter
M.
Umbarger
H.E.
, Eds.), pp.
542
560
.
American Society for Microbiology
,
Washington, DC
.
[4]
Plamann
M.D.
Stauffer
G.V.
(
1989
)
Regulation of the Escherichia coli glyA gene by the metR gene product and homocysteine
.
J. Bacteriol.
 
171
,
4958
4962
.
[5]
Lorenz
E.
Stauffer
G.V.
(
1995
)
Characterization of the MetR binding sites for the glyA gene in Escherichia coli
.
J. Bacteriol.
 
177
,
4113
4120
.
[6]
Vogel
H.J.
Bonner
D.M.
(
1956
)
Acetylornithase of Escherichia coli: partial purification and some properties
.
J. Biol. Chem.
 
218
,
97
106
.
[7]
Stauffer
G.V.
Brenchley
J.E.
(
1977
)
Influence of methionine biosynthesis on serine transhydroxymethylase regulation in Salmonella typhimurium
.
J. Bacteriol.
 
129
,
740
749
.
[8]
Maniatis
T.
Fritsch
E.F.
Sambrook
J.
(
1982
)
Molecular Cloning: A Laboratory Manual
 .
Cold Spring Harbor Laboratory
,
Cold Spring Harbor, NY
.
[9]
Sanger
F.M.
Nicklen
S.
Coulson
A.R.
(
1977
)
DNA sequencing with chain-terminating inhibitors
.
Proc. Natl. Acad. Sci. USA
 
74
,
5463
5467
.
[10]
Sankar
G.
Sommer
S.S.
(
1990
)
The ‘Megaprimer’ method of site-directed mutagenesis
.
BioTechniques
 
8
,
404
407
.
[11]
Casadaban
M.J.
Chou
J.
Cohen
S.N.
(
1980
)
In vitro gene fusions that join an enzymatically active β-galactosidase segment to ammo-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals
.
J. Bacteriol.
 
143
,
971
980
.
[12]
Urbanowski
M.L.
Stauffer
G.V.
(
1986
)
Autoregulation by tandem promoters of the Salmonella typhimurium LT2 met J gene
.
J. Bacteriol.
 
165
,
740
745
.
[13]
Shimada
K.
Weisberg
R.A.
Gottesmann
M.E.
(
1972
)
Prophage lambda at unusual chromosomal locations. I. location of the secondary attachment sites and the properties of the lysogens.
.
J. Mol. Biol.
 
63
,
483
503
.
[14]
Lorenz
E.
Plamann
M.D.
Stauffer
G.V.
(
1996
)
Escherichia coli mutants with increased glyA gene expression
.
Mol. Gen. Genet.
 
250
,
81
88
.
[15]
Urbanowski
M.L.
Stauffer
G.V.
(
1987
)
Regulation of the metR gene of Salmonella typhimurium
.
J. Bacteriol.
 
169
,
5841
5844
.
[16]
Lorenz
E.
Stauffer
G.V.
(
1996
)
Cooperative MetR binding in the Escherichia coli glyA control region
.
FEMS Microbiol. Lett.
 
137
,
147
152
.
[17]
Plamann
M.D.
Stauffer
G.V.
(
1983
)
Characterization of the Escherichia coli gene for serine hydroxymethyltransferase
.
Gene
 
22
,
9
18
.
[18]
Steiert
J.G.
Rolfes
R.J.
Zalkin
H.
Stauffer
G.V.
(
1990
)
Regulation of the Escherichia coli glyA gene by the pur R gene product
.
J. Bacteriol.
 
172
,
3799
3803
.
[19]
Miller
J.H.
(
1992
)
A Short Course in Bacterial Genetics
 .
Cold Spring Harbor Laboratory
,
Cold Spring Harbor, NY
.