The relA locus and the regulation of lysine biosynthesis in Escherichia coli

Patte, Jean‐Claude; Morand, Philippe; Boy, Emmanuelle; Richaud, Catherine; Borne, Françoise

doi:10.1007/bf00425459

Cited by 22 publications

(10 citation statements)

References 42 publications

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“…The larger fusion (strain JCP75) behaved as the fusion carried by a multicopy plasmid and showed a fourfold reduction in ␤-galactosidase synthesis in the presence of lysine (5). Only chemostat-induced lysine limitation can lead to a 10-fold derepression of the dapB gene (20). It also exhibited a twofold reduction in the presence of arginine.…”

Section: Resultsmentioning

confidence: 98%

Lysine Represses Transcription of the Escherichia coli dapB Gene by Preventing Its Activation by the ArgP Activator

et al. 2008

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The Escherichia coli dapB gene encodes one of the enzymes of the biosynthetic pathway leading to lysine and its immediate precursor, diaminopimelate. Expression of dapB is repressed by lysine, but no trans-acting regulator has been identified so far. Our analysis of the dapB regulatory region shows that sequences located in the ؊81/؊118 interval upstream of the transcription start site are essential for full expression of dapB, as well as for lysine repression. Screening a genomic library for a gene that could alleviate lysine repression when present in multicopy led to the recovery of argP, a gene encoding an activating protein of the LysR-type family, known to use lysine as an effector. An argP null mutation strongly decreases dapB transcription that becomes insensitive to lysine. Purified His 6 -tagged ArgP protein binds with an apparent K d of 35 nM to the dapB promoter in a gel retardation assay, provided that sequences up to ؊103 are present. In the presence of L-lysine and L-arginine, the binding of ArgP to dapB is partly relieved. These results fit with a model in which ArgP contributes to enhanced transcription of dapB when lysine becomes limiting.

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Section: Resultsmentioning

confidence: 98%

Lysine Represses Transcription of the Escherichia coli dapB Gene by Preventing Its Activation by the ArgP Activator

et al. 2008

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“…E. coli strains used in this work are MC4100 (araD139, AlacU169, rpsL, thi, relA) (12) and its isogeneic derivatives RM4102 (relA+) (13) and RDB22 (relA', dapB) (2). Cultures were grown in minimal medium 63 (14) supplemented with 0.5% glucose and, when specified, with L-arginine (100 Ag/ml) or uracil (50 jxg/ml) plus cytidine (100 jig/ml).…”

Section: Methodsmentioning

confidence: 99%

Multiple regulatory signals in the control region of the Escherichia coli carAB operon.

Bouvier¹,

Patte²,

Stragier³

1984

Proc. Natl. Acad. Sci. U.S.A.

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The first reaction in pyrimidine and arginine biosynthesis in Escherichia coli is catalyzed by a single enzyme, carbamoyl-phosphate synthetase (EC 6.3.5.5), the product of the carAB operon. Expression of this operon is cumulatively repressed by arginine and pyrimidines. The nucleotide sequence of the carAB control region was determined and transcriptional starts were localized. Two adjacent promoters, 70 base pairs apart, appear to be used in vivo, the downstream one overlapping a typical arginine operator. The absence of any attenuation-like sequence excludes such a mechanism for pyrimidine-mediated repression. Various fragments of the carA promoter-proximal region were fused in vitro with the lacZ gene. Results obtained with these fusions indicate that (i) translation of the carA gene can be initiated in vivo without an AUG codon but very likely with an UUG or an AUU codon; (ii) the carAB downstream promoter is repressed by arginine; and (iii) the carAB upstream promoter is repressed by pyrimidines and subject to stringent control. When carried by a multicopy plasmid the carAB control region escapes repression by arginine and pyrimidines. The existence of a pyrimidine repressor, present in limiting amounts in the cell, is therefore postulated.A X bacteriophage carrying the Escherichia coli dapB gene, one of eight loci involved in lysine biosynthesis (1), has been isolated in our'laboratory (2). After transfer to a plasmid, the dapB gene was subcloned as a 2.3-kilobase fragment and completely sequenced (unpublished data). Downstream of the dapB coding sequence a large open reading frame was found, preceded by strong putative transcriptional signals. Using another dapB transducing phage, Mackie (3) had independently shown that this region codes for a 48,000-dalton polypeptide, which he subsequently proposed (4) to be the product of the carA gene, the closest locus on the E. coli genetic linkage map (5). This gene is part of the carAB operon (6); it encodes the light subunit of carbamoyl-phosphate synthetase (glutamine hydrolyzing) (EC 6.3.5.5), an enzyme catalyzing the synthesis of carbamoyl phosphate, the common precursor of pyrimidines and arginine (7). Its expression is cumulatively repressed by arginine and pyrimidines (8) through a transcriptional mechanism (9) involving the arginine repressor (10). Some of our data were consistent with Mackie's hypothesis, which was fully confirmed by unpublished work from N. Glansdorff and A. Pierard's laboratory (see ref. 11). This paper describes experiments performed to explain the molecular basis of the carAB operon double regulation.MATERIALS AND METHODS Bacterial Strains and Media. E. coli strains used in this work are MC4100 (araD139, AlacU169, rpsL, thi, relA) (12) and its isogeneic derivatives RM4102 (relA+) (13) and RDB22 (relA', dapB) (2). Cultures were grown in minimal medium 63 (14) supplemented with 0.5% glucose and, when specified, with L-arginine (100 Ag/ml) or uracil (50 jxg/ml) plus cytidine (100 jig/ml). Isoleucine starvation was induced by addi...

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“…It can be seen that: (i) in wildtype strain RM 4102 with plasmid pAD20, ASA-dehydrogenase specific activity is increased -15-fold (line 1); (ii) the presence of excess lysine, threonine, and methionine during ©g IRL Press Limited, Oxford, England. 0261-4189/82/0103-0379$2.00/0 379 lln dIII ta N lindl hl II (Patte et al, 1980). bRM 4104 same markers as RM 4102, lysA 1101 (Patte et al, 1980).…”

Section: Isolation and Purification Of The Asd Genementioning

confidence: 99%

“…The asd gene encodes aspartic semialdehyde dehydrogenase (ASA-dehydrogenase; EC 1.2.1.11). This gene is interesting for several reasons: (i) it belongs to the common part of the biosynthetic pathways of lysine, threonine, and methionine (Theze et al, 1974); (ii) its synthesis is multivalently repressed by these amino acids, the efficiency of derepression being greatest in the case of lysine limitation (Boy and Patte, 1972); (iii) a regulatory (and unexplained) role of glucose-6-phosphate has also been described, possibly at the transcriptional level (Boy and Patte, 1979); (iv) the allelic state of the relA gene modifies the expression of the gene (Patte et al, 1980); and (v) the protein has been purified; it is a dimer composed of identical protomers of mol. wt.…”

Section: Introductionmentioning

confidence: 99%

Nucleotide sequence of the asd gene of Escherichia coli: absence of a typical attenuation signal.

Haziza¹,

Stragier²,

Patte³

1982

The EMBO Journal

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The asd gene of escherichia coli encodes aspartic semialdehyde dehydrogenase, an enzyme involved in lysine, threonine, and methionine biosynthesis; its synthesis is controlled by a multivalent repression mechanism. It was cloned in plasmid pBR322 and its complete nucleotide sequence determined. The sequence predicts a polypeptide chain of 367 amino acids, in good agreement with results obtained for the purified protein (Biellmann et al., 1980a). Our data indicate a Cys residue instead of a His residue, which was proposed after covalent labeling of the active center of the enzyme; this is more in line with the catalytic site of glyceraldehyde‐3‐phosphate dehydrogenase, an enzyme which carries out a similar reaction. The nucleotide sequence that precedes the translational start does not display any of the characteristic features of an attenuation signal. Hence the expression of the asd gene is probably not controlled in the same way as other multivalently repressed operons such as ilva and thr.

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The relA locus and the regulation of lysine biosynthesis in Escherichia coli

Cited by 22 publications

References 42 publications

Lysine Represses Transcription of the Escherichia coli dapB Gene by Preventing Its Activation by the ArgP Activator

Lysine Represses Transcription of the Escherichia coli dapB Gene by Preventing Its Activation by the ArgP Activator

Multiple regulatory signals in the control region of the Escherichia coli carAB operon.

Nucleotide sequence of the asd gene of Escherichia coli: absence of a typical attenuation signal.

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